Wound and skin care compositions

ABSTRACT

The invention provides compositions and methods that employ compounds that can stimulate proliferation of fibroblasts or keratinocytes and/or stimulate production of collagen by fibroblasts. These compositions and methods are useful for treating gum- and skin-related conditions.

FIELD OF THE INVENTION

[0001] The present invention relates generally to the field of woundhealing and to the repair and maintenance of healthy skin.

BACKGROUND OF THE INVENTION

[0002] Skin is subject to insults by many extrinsic and intrinsicfactors. Extrinsic factors that can adversely affect the skin includeultraviolet radiation (e.g., from sun exposure), environmentalpollution, wind, heat, low humidity, harsh surfactants, abrasives, andthe like. Intrinsic factors that lead to skin problems includechronological aging and other biochemical changes from within the skin.Whether extrinsic or intrinsic, these factors result in visible signs ofskin aging and damage, such as wrinkling, roughness and histologicalchanges. Moreover, to many people, skin wrinkles are a reminder of thedisappearance of youth. As a result, the elimination of wrinkles hasbecome a concern for many people. Anti-wrinkle treatments range fromcosmetic creams and moisturizers to various forms of cosmetic surgery.

[0003] Extrinsic or intrinsic factors may result in the thinning andgeneral degradation of the skin. For example, as the skin naturallyages, there is a reduction in the cells and the blood vessels thatsupply the skin. There is also a flattening of the dermal-epidermaljunction that results in weaker mechanical resistance of this junction.See, for example, Oikarinen, “The Aging of Skin: Chronoaging VersusPhotoaging,” Photodermatol. Photoimmunol. Photomed., vol. 7, pp. 3-4,1990, which is incorporated by reference herein in its entirety.

[0004] Skin contains an elaborate network of elastin fibers that isresponsible for maintaining its elastic properties. With excessiveexposure to sunlight the elastic fiber system becomes hyperplastic,disorganized and ultimately disrupted. This process is known as actinicelastosis and it is a principal cause of wrinkling, discoloration andlaxity of the skin in the exposed areas of the body. As new fibroblasts,endothelial cells and keratinocytes form, the skin can repair itself.However, the skin becomes less able to do so as it ages. Therefore,agents that can accelerate the growth and repair of prematurely agedskin are needed.

[0005] Wound healing is also accelerated by increased cellularproliferation and migration of certain cell types. The mechanismsinvolved in wound healing are often divided into four phases:hemostasis, inflammation, proliferation and maturation. Duringinflammation, leucocytes accumulate to combat bacteria and thepermeability blood vessel walls increases, leading to swelling. If aninfection does not develop the number of leucocytes diminishes.Monocytes replace the leukocytes. Macrophages and lymphocytes releasegrowth factors (cytokines) as well as a number of chemical substances,such as histamine, serotonin, and prostaglandins. These substances helpregulate the wound healing process. In the proliferation phase, newfibroblasts, endothelial cells and keratinocytes arise, connectivetissue is formed, new blood vessels grow and injured tissue isregenerated. Fibroblasts become dominant after about a week, theinflammation decreases and the strength of the tissues around the woundsite increases rapidly. During the maturation phase collagen is laiddown and scar tissue is formed. This maturation phase might go on for along time during which time, tissues of various types are regenerated.In order to obtain an optimal healing of skin and associated tissues,the supply of different vitamins and trace elements as well as nutrientsshould be sufficient as well as the oxygen supply.

[0006] While certain skin care compositions are available on the market,such compositions do not effectively stimulate the growth of new skintissues. Hence, there is a need for formulations that not only improvethe appearance but also the health of skin. Ideally, such formulationswould provide a range of useful activities such as, for example, woundhealing, scar reduction, soothing rashes, eliminating wrinkles andreducing the signs of aging.

SUMMARY OF THE INVENTION

[0007] The invention provides compositions that can stimulate theformation of new skin and gum tissues, facilitate wound healing, andameliorate the effects of aging. In particular, the compositions andmethods of the invention can stimulate cellular proliferation ofmammalian fibroblasts or keratinocytes. Not only can these compositionsstimulate cell growth and the repair of skin and gum tissues, but theycan also stimulate collagen production in mammalian fibroblasts.

[0008] Thus, the invention provides compositions comprising adermatologically or pharmaceutically acceptable carrier and an effectiveamount of a jasmonic acid compound, a gibberellic acid compound or azeatin compound; wherein the composition can stimulate cellularproliferation of mammalian fibroblasts or keratinocytes. Suchcompositions are useful for treating wounds, stimulating gum tissuegrowth and for reversing the effects of aging on skin.

[0009] In other embodiments, the invention provides a compositioncomprising a dermatologically or pharmaceutically acceptable carrier andan effective amount of a jasmonic acid compound, a gibberellic acidcompound or a zeatin compound; wherein the composition can stimulatecollagen production in mammalian fibroblasts. Such compositions areuseful for treating wounds, stimulating gum tissue growth and forreversing the effects of aging on skin.

[0010] Further embodiments of the invention relate to compositionscomprise a dermatologically or pharmaceutically acceptable carrier andan effective amount of a hydroxy acid compound, wherein the compositioncan stimulate cellular proliferation of mammalian fibroblasts orkeratinocytes. Such compositions can further comprise an effectiveamount of a jasmonic acid compound, a gibberellic acid compound or azeatin compound.

[0011] The present invention also provides an orally acceptableformulation for the treatment of gum tissues comprising an acceptablecarrier and an effective amount of a jasmonic acid compound, agibberellic acid compound or a zeatin compound; wherein the formulationcan stimulate cellular proliferation of mammalian fibroblasts orkeratinocytes. The composition can stimulate collagen production inmammalian fibroblasts.

[0012] In one embodiment, the invention provides a compositioncomprising:

[0013] a) from about 0.01% to about 30%, by weight of the composition,of jasmonic acid, gibberellic acid or zeatin;

[0014] b) from about 0.01% to about 30%, by weight of the composition,of acetylsalicylic acid or salicylic acid; and

[0015] c) a dermatologically or pharmaceutically acceptable carrier.

[0016] The invention also provides a method of stimulating growth offibroblasts or keratinocytes comprising administering to the fibroblastsor the keratinocytes a safe and effective amount of a compositioncomprising a dermatologically acceptable carrier and an effective amountof a hydroxy acid compound, wherein the composition can stimulatecellular proliferation of mammalian fibroblasts or keratinocytes.

[0017] In another embodiment, the invention provides a method ofstimulating growth of fibroblasts or keratinocytes comprisingadministering to the fibroblasts or the keratinocytes a safe andeffective amount of a composition comprising a dermatologicallyacceptable carrier and an effective amount of a jasmonic acid compound,a gibberellic acid compound or a zeatin compound; wherein thecomposition can stimulate cellular proliferation of mammalianfibroblasts or keratinocytes.

[0018] The invention also provides a method of stimulating collagenproduction in mammalian fibroblasts comprising administering to thefibroblasts a safe and effective amount of a composition comprising adermatologically acceptable carrier and an effective amount of ajasmonic acid compound, a gibberellic acid compound or a zeatincompound; wherein the composition can stimulate collagen production inmammalian fibroblasts. The composition can further comprise a hydroxyacid compound.

[0019] In some embodiments, the effective amount of the jasmonic acidcompound, the gibberellic acid compound or the zeatin compound is aconcentration of about 0.001 micromolar to about 10 millimolar. In otherembodiments, the effective amount of the jasmonic acid compound, thegibberellic acid compound or the zeatin compound is a concentration ofabout 1 micromolar to about 5 millimolar.

[0020] The compositions can have one or more additional ingredients. Forexample, the compositions can have additional ingredients such asdesquamation compounds, anti-acne compounds, anti-wrinkle compounds,vitamin B₃ compounds, vitamin E compounds, retinoid compounds, hydroxyacid compounds, anti-oxidant compounds, radical scavengers, chelatingagents, flavonoid compounds, anti-inflammatory agents, anti-celluliteagents, topical anesthetics, tanning compounds, skin lightening agents,skin healing compounds, anti-microbial compounds, antifungal compounds,sunscreen compounds, particulate material, moisturizers, or thickeningagents.

[0021] Examples of vitamin E compounds that can be used includetocopherol, tocopherol acetate, a tocopherol ester or a mixture thereof.Examples of retinoid compounds that can be used include retinol,retinal, retinol ester, retinyl propionate, retinoic acid, retinylpalmitate, or a mixture thereof. Particulate materials that may beemployed include mica, mica treated with barium sulfate and TiO₂,silica, nylon, polyethylene, talc, styrene, polypropylene,ethylene/acrylic acid copolymer, sericite, aluminum oxide, siliconeresin, barium sulfate, titanium dioxide, iron oxide, bismuthoxychloride, calcium carbonate, cellulose acetate, polymethylmethacrylate, or a mixture thereof. Sunscreens that can be used include,for example, a metallic oxide selected from the group consisting oftitanium dioxide having an average primary particle size of from about15 nm to about 100 nm, zinc oxide having an average primary particlesize of from about 15 nm to about 150 nm, zirconium oxide having anaverage primary particle size of from about 15 nm to about 150 nm, ironoxide having an average primary particle size of from about 15 nm toabout 500 nm, or a mixture thereof. Other sunscreens that may be usedinclude octylmethoxycinnamate, octyl salicylate, terephthalyidenedicamphor sulfonic acid, avobenzone, octocrylene, or a mixture thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0022]FIG. 1 illustrates the cell proliferating activity ofacetylsalicylic acid on human skin fibroblasts. The optical density (OD)at 490 nm was used as a measure of cellular density. The concentrationof acetylsalicylic acid tested varied between 1×10⁻⁴ M (designatedASA4), 1×10⁻⁵ M (designated ASA5) and 1×10⁻⁶ M (designated ASA6).Control cells received no acetylsalicylic acid. As illustrated, allconcentrations of acetylsalicylic acid had a significant effect on cellgrowth (P<0.001). However, the effect of acetylsalicylic acid was dosedependent, with a greater effect observed at lower concentrations.

[0023]FIG. 2 illustrates the cell proliferating activity of salicylicacid on human skin fibroblasts. The optical density (OD) at 490 nm wasused as a measure of cellular density. The concentration of salicylicacid tested varied between 1×10⁻⁴ M (designated SA4), 1×10⁻⁵ M(designated SA5) and 1×10⁻⁶ M (designated SA6). Control cells receivedno salicylic acid. As illustrated, all concentrations of salicylic acidhad a significant effect on cell growth (P<0.01 to P<0.001), but lowerconcentrations of salicylic acid (1×10⁻⁵ M and 1×10⁻⁶ M) had a greatereffect (P<0.001).

[0024]FIG. 3 illustrates the cell proliferating activity of jasmonicacid on human skin fibroblasts. The optical density (OD) at 490 nm wasused as a measure of cellular density. The concentration of jasmonicacid tested varied between 1×10⁻⁴ M (designated JA4), 1×10⁻⁵ M(designated JA5) and 1×10⁻⁶ M (designated JA6). Control cells receivedno jasmonic acid. As illustrated, all concentrations of jasmonic acidhad an effect on cell growth (P<0.05 to P<0.001), but lowerconcentrations of jasmonic acid (1×10⁻⁵ M and 1×10⁻⁶ M) had a greatereffect (P<0.001).

[0025]FIG. 4 illustrates the cell proliferating activity of t-zeatin onhuman skin fibroblasts. The optical density (OD) at 490 nm was used as ameasure of cellular density. The concentration of t-zeatin tested variedbetween 1×10⁻⁴ M (designated ZA4), 1×10⁻⁵ M (designated ZA5) and 1×10⁻⁶M (designated ZA6). Control cells received no t-zeatin. As illustrated,all concentrations of t-zeatin had a significant effect on cell growth(P<0.001). However, the effect of zeatin was dose dependent, with agreater effect observed at lower concentrations.

[0026]FIG. 5 illustrates the cell proliferating activity of gibberellicacid on human keratinocytes. The optical density (OD) at 490 nm was usedas a measure of cellular density. The concentration of gibberellic acidtested varied between 1×10⁻⁴ M (designated GA4), 1×10⁻⁵ M (designatedGA5) and 1×10⁻⁶ M (designated GA6). Control cells received nogibberellic acid. As illustrated, all concentrations of gibberellic acidhad a significant effect on cell growth (P<0.001). However, the effectof gibberellic acid was dose dependent, with a greater effect observedat lower concentrations.

[0027]FIG. 6 illustrates the effect of jasmonic acid (JA), gibberellicacid (GA) and zeatin (ZA) on collagen production by human fibroblastcells. The optical density (OD) at 450 nm was used as an indicator ofcollagen production. The concentration of compound tested varied between1×10⁻⁴ M (designated 4), 1×10⁻⁵ M (designated 5) and 1×10⁻⁶ M(designated 6). Control cells received no jasmonic acid, gibberellicacid or zeatin. As illustrated, all compounds stimulated collagenproduction but lower concentrations of gibberellic acid had the mostprofound effect. Higher concentrations of jasmonic acid and mediumconcentrations of zeatin also stimulated collagen production.

DETAILED DESCRIPTION OF THE INVENTION

[0028] The invention provides compositions for increasing cellproliferation and for stimulating collagen production in keratinoustissues such as the skin. These compositions can be used to reverse theeffects of aging. Such compositions are useful for treating wounds,stimulating gum tissue growth and for reversing the effects of aging onskin. Such compositions are useful for treating wounds, stimulating gumtissue growth and for reversing the effects of aging on skin. Suchcompositions are useful for treating wounds, stimulating gum tissuegrowth and for reversing the effects of aging on skin. Such compositionsare useful for treating wounds, stimulating gum tissue growth and forreversing the effects of aging on skin. The compositions of theinvention can contain an effective amount of a hydroxy acid compound, ajasmonic acid compound, a gibberellic acid compound or a zeatin compoundas well as acceptable carriers and other ingredients.

[0029] Definitions

[0030] The term “keratinous tissue,” as used herein, refers tokeratin-containing layers disposed as the outermost protective coveringof mammals (e.g., humans, dogs, cats, etc.) that includes, but is notlimited to, skin, lips, hair, toenails, fingernails, cuticles, hooves,etc.

[0031] The term “topical application”, as used herein, means to apply orspread the compositions of the present invention onto the surface of thekeratinous tissue.

[0032] The term “dermatologically-acceptable,” as used herein, meansthat the described compositions or components thereof are suitable foruse in contact with mammalian keratinous tissue without undue toxicity,incompatibility, instability, allergic response, and the like.

[0033] The term “safe and effective amount” as used herein means anamount of a compound or composition sufficient to significantly induce apositive benefit, such as a positive keratinous tissue appearance orfeel benefit, including independently or in combinations the benefitsdisclosed herein, but low enough to avoid serious side effects, i.e., toprovide a reasonable benefit to risk ratio, within the scope of soundjudgment of the skilled artisan.

[0034] The term “sagging” as used herein means the laxity, slackness, orthe like condition of skin that occurs as a result of loss of, damageto, alterations to, and/or abnormalities in dermal elastin.

[0035] The terms “smoothing” and “softening” as used herein meanaltering the surface of the keratinous tissue such that its tactile feelis improved. “Signs of skin aging” include, but are not limited to, alloutwardly visible or tactilely perceptible manifestations as well as anyother macro or micro effects due to skin aging. Such signs may beinduced or caused by intrinsic factors or extrinsic factors, e.g.,chronological aging and/or environmental damage. These signs may resultfrom processes that include, but are not limited to, the development oftextural discontinuities such as wrinkles and coarse deep wrinkles, skinlines, crevices, bumps, large pores (e.g., associated with adnexalstructures such as sweat gland ducts, sebaceous glands, or hairfollicles), or unevenness or roughness, loss of skin elasticity (lossand/or inactivation of functional skin elastin), sagging (includingpuffiness in the eye area and jowls), loss of skin firmness, loss ofskin tightness, loss of skin recoil from deformation, discoloration(including under eye circles), blotching, sallowness, hyperpigmentedskin regions such as age spots and freckles, keratoses, abnormaldifferentiation, hyperkeratinization, elastosis, collagen breakdown, andother histological changes in the stratum corneum, dermis, epidermis,the skin vascular system (e.g., telangiectasia or spider vessels), andunderlying tissues, especially those proximate to the skin.

[0036] Effects of Aging on Skin

[0037] Skin, which is the external covering of the body, has twocomponents: the epidermis that contains four layers and the dermis, alsoreferred to as the corium, cutis, derma, or true skin, that contains asuperficial papillary layer and a deep reticular layer. Collagenconstitutes about 80% of the dry weight of the dermis and is the majorfibrillar component of human skin. The dermis is composed of connectivetissue that contains lymphatics, nerves and nerve endings, bloodvessels, sebaceous and sweat glands, and elastic fibers that provide theelastic properties of the skin. The mature fiber contains about 90%elastin and two glycosaminoglycans are present at concentrations ofabout 2% and 0.1%, respectively (see, e.g., Braverman (1982) J. Invest.Dermatol, 78:434-443). The coarse branching fibers are entwined withcollagen fiber bundles in the reticular dermis. The fibers rise from thedeeper layers of the papillary layer of the dermis and, as they risetowards the epidermis, they split repeatedly become finer and form anetwork.

[0038] As humans age, there are changes in the quantity and integrity ofdermal elastic tissue (Warren et al. (1991) J. of the American Acad.Dermatology 25:751-760). Gross alteration in elastin leads toalterations in the appearance of the skin (see, Bryce et al. (1988) J.Invest. Dermatology 91:175-180; Kornberg et al. (1985) New Engl. J. Med.312:771-774; Shelley et al. (1977) Br. J. Dermatol, 7:441-445). Theassociation of changes in the fibers and the appearance of wrinklesindicate a causal relationship between the integrity of the elasticfiber network and the mechanical properties of the skin. Aging skin ischaracterized by initial elastogenesis followed by a slow spontaneousprogressive degradation of the elastic fibers that leads to laxity andwrinkling. Studies of skin from subjects of various ages indicate thatthe degradation of elastic fibers that begins about age 30 and becomesmarked after age 70 is a major feature of aging skin.

[0039] In understanding the process of aging of human skin, it ispertinent to understand the role of fibroblasts, particularly in thedermis of human skin and in the corresponding connective tissue layerunderlying the integuments of other mammals and the epithelia of theinner wall of the gastrointestinal tracts of humans and other animals.Fibroblasts synthesize components that are required for maintenance ofthe structural, functional and cosmetic integrity of the skin and thestructural and functional integrity of other surface tissues covered byepithelia. These components include collagen and elastin, which arefibrous proteins responsible for the three-dimensional architecture ofskin and the other surface tissues, fibronectin, which is a proteinresponsible for cell anchorage and maintenance of cell morphology; and anumber of proteinaceous growth factors essential for the maintenance ofepithelia and basal cell layers and connective tissue layers underlyingthem. Available evidence indicates that protein biosynthetic activity offibroblasts decreases significantly with age. For example, the rate ofcollagen synthesis at about 70% of life expectancy for human fibroblastsin culture is only about 50% of that of such fibroblasts at less than20% of life expectancy.

[0040] The changes in the appearance of the skin with age result fromnatural or intrinsic aging superimposed by actinic damage resulting fromphotoaging (see, e.g., Weiss et el. (1988) J. American Medical Assoc.259:527-532). Intrinsic aging includes changes that occur as a result ofendogenous factors and genetically programmed senescence, includingepidermal and dermal atrophy. Photoaging results from long-term exposureto UV (ultra-violet) radiation, primarily from the sun. Ultravioletexposure is also associated with tumor induction and other skinpathologies. Ultraviolet radiation from the sun includes UVB (280-315nm) and the more penetrating UVA (315-400 nm) radiation. UVB causeserythema, skin cancer and dermal connective tissue damage. UVA alsocauses erythema and is carcinogenic at higher doses. Low doses (2.5minimal erythemic doses (MEDs)) are sufficient to cause endothelial cellenlargement, extravasation of blood cells, and perivenular neutrophilinfiltrates as wells increased concentrations of mediators of theinflammatory response (see, e.g., Kligman et al. (1985) J. Invest.Dermatol. 84:272-276).

[0041] Long-term ultraviolet exposure results in histological andvisible changes in the skin, including: damage to the underlyingconnective tissue, manifested as elastosis and increases in theglycosaminoglycans and loss of collagen; dermal accumulation ofelastin-staining material resulting from the degenerative changes incollagen fibers; epidermal dysplasia with cytologic atypia and loss ofpolarity of keratinocytes; and an inflammatory infiltrate (see, e.g.,Bissett et al. (1987) Photochemistry and Microbiology 46:367-378). Thedegradation of elastic fibers and wrinkling associated withintrinsically aging skin also accompanies photoaging. In humans,advanced photodamage can be detected in the staining properties ofdermal tissue resulting from changes in the insoluble and solublefractions of collagen that occur as the entire upper dermis becomesfilled with elastosis (Kligman et al. (1989) J. Investigative Dermatol.93:210-214). The changes in collagen and elastic fiber over decades ofsuch exposure result in skin that is wrinkled, yellowed, blotchy, lax,rough and leathery. Scanning electron microscopy of aged skin indicatesthat the network of elastic fibers becomes denser and has a moredisorganized arrangement than younger skin.

[0042] Exposure to sunlight is such a pronounced factor in prematureaging that by middle age individuals who have been exposed to moresunlight appear older than those who have not. The extent of dermaldegenerative change correlates with the visible signs of prematureaging. The subepidermal band of normal dermis, which is a site ofcontinual dermal repair, contains normal collagen fibers. This zonebecomes visually evident, however, only after there is sufficientelastotic damage to delineate this region. The elastotic material iscomposed principally of elastin and microfibrillar proteins thatcodistribute with fibronectin (see, Schwartz (1988) J. Invest. Dermatol.1:158-161). Actinic elastosis appears to be reversible to some extent bytreatment with chemical peels, dermabrasion or topical application oftretinoin (see, e.g., Warren et al. (1991) J. American Acad. Dermatol.25:751-760; and Weiss et al. (1988) J. American Medical Assoc.259:527-532).

[0043] Hydroxy Acids

[0044] The compositions of the present invention may contain a safe andeffective amount of a hydroxy acid. The hydroxy acids can bealpha-hydroxy or beta-hydroxy acids. Examples of hydroxy acids for usein the compositions of the present invention include salicylic acid,lactic acid, glycolic acid, acetylsalicylic acid, and other salicylicacid derivatives. In some embodiments, salicylic acid and the syntheticand naturally occurring derivatives of salicylic acid can be used in thecompositions and methods of the present invention.

[0045] wherein:

[0046] R1 is COOR, or —(CH₂)n-OX

[0047] R is H, or alkyl (e.g., with one to twenty carbons);

[0048] n is an integer of about 1 to about 20;

[0049] X is H, or 1 to 6 sugar residues (e.g., hexoses or pentoses);

[0050] R2 is COOR, —(CH₂)n-OX, OCO-alkyl (C1-C20), OY; and

[0051] Y is H, alkyl (C1-20), or 1 to 6 sugar residues (e.g., hexoses orpentoses).

[0052] In general, the alkyl groups employed in these hydroxy acidcompounds have about one to twenty carbon atoms, although in someembodiments lower alkyl groups are used, for example, alkyl groups withabout one to eight carbon atoms. Alkyl groups with even lower numbers ofcarbon atoms can also be used, for example, alkyl groups with one tosix, or one to three carbon atoms.

[0053] In some embodiments, salicylic acid is employed in thecompositions of the invention. Salicylic acid is a compound of formula Iwherein R1 is COOH and R2 is OH. In other embodiments, acetylsalicylicacid is employed in the compositions of the invention. Acetylsalicylicacid is a compound of formula I wherein R1is COOH and R2 is OCOCH₃.

[0054] When present in the compositions of the present invention,salicylic acid or its derivatives can be used in an amount of from about0.001% to about 50%, or from about 0.01% to about 20%, or from about0.01% to about 10%, or from about 0.05% to about 5%, or from about 0.05%to about 2% of the composition. According to the invention, in situconcentrations of acetylsalicylic acid or salicylic acid that range fromabout 10⁻⁴ M to about 10⁻⁶ M are effective for increasing cellproliferation and stimulating the production of collagen in mammaliankeratinous tissue.

[0055] Gibberellic Acid

[0056] Gibberellic acid comprises a class of compounds that is alsoreferred to as gibberellins. Gibberellins are plant hormones that affecta wide variety of processes throughout the life cycle of plants,including seed germination, stem elongation, flower induction, antherdevelopment, and seed and pericarp growth. Gibberellins are tetracyclicditerpenoid acids found in fungi and higher plants having theent-gibberellane ring system shown in the following structure (II).

[0057] Gibberellins were first isolated by Japanese researchers in the1930s from cultures of the fungus Gibberella fujikuroi (Fusariummoniliforme). These secondary metabolites have been shown to be presentin other fungal species, in some ferns, and in many gymnosperms andangiosperms. Of the 121 known gibberellins, 96 have been identified onlyin higher plants, 12 are present only in Gibberella, and 12 are presentin both. As in Gibberella, many different gibberellins can be present inindividual angiosperms.

[0058] Two main types of gibberellins exist: the C20-gibberellins, whichhave 20 carbon atoms (structure III, below), and the C19-gibberellins,in which the twentieth carbon atom has been lost due to metabolism(structure IV, below). The carboxylic acid at carbon-19 bonds tocarbon-10 to produce a lactone bridge in almost all of theC₁₉-gibberellins.

[0059] The ent-gibberellane ring system can contain many structuralmodifications, accounting for the large number of known gibberellins.Naturally occurring gibberellins with identified structures areallocated an “A number” (MacMillan et al. (1968) Nature 217:170-171). Atpresent, 126 naturally occurring gibberellins of plant and fungal originare known. Current structural information on gibberellins can be foundat the website plant-hormones.bbsrc.ac.uk/gibberellin_information2.htm.

[0060] Variations in gibberellin structure arise in several ways.Carbon-20 can exist in different oxidative states, e.g., methyl (—CH₃),hydroxymethyl (—CH₂OH), aldehyde (—CHO), or carboxylic acid (—COOH). Theent-gibberellane skeleton, especially that of the C19-gibberellins, canalso contain additional functional groups. Hydroxyl (—OH) groups arefrequently inserted into the ring system; insertion of epoxide (>O) andketone (═O) functions also occurs, although less commonly. The positionand/or stereochemistry of substituent groups can affect the biochemicaland physiological significance of the molecules. Substituent groupspositioned above the ring plane are said to be in the β-configuration;their bonding to the ring is designated by a solid, elongated triangle.Substituent groups positioned below the ring plane are said to be in theα-configuration; their bonding to the ring is designated by a dashed,elongated triangle. The attachment of substituent groups in the plane ofthe ring system is indicated by a straight line.

[0061] Gibberellins can exist as conjugates, for example with a moleculeof glucose, either by an ether or an ester linkage. Such conjugation maytemporarily or permanently inactivate a gibberellin.

[0062] The biological activity of different gibberellins varies, andvarious gibberellins within a plant can be precursors, biosyntheticintermediates, or deactivation products of active gibberellins. Threestructural features are commonly associated with gibberellin biologicalactivity: a 3-hydroxyl group, a 7-carboxyl group, and a lactone ring.Broadly speaking, a compound possessing the ent-gibberellane ring systembut lacking one or more of these structural features can be considered agibberellin precursor, an intermediate, or a derivative.

[0063] The compositions and methods of the invention generally employactive forms of gibberellic acids, for example, those having structuresrelated to the following structure (V).

[0064] When present in the compositions of the present invention,gibberellic acids or their derivatives can be used in an amount of fromabout 0.001% to about 50%, or from about 0.01% to about 20%, or fromabout 0.01% to about 10%, or from about 0.05% to about 5%, or from about0.05% to about 2% of the composition. According to the invention, insitu concentrations of an active gibberellic acid ranging from about10⁻⁴ M to about 10⁻⁶ M are effective for increasing cell proliferationand stimulating the production of collagen in mammalian keratinoustissue.

[0065] Jasmonic Acid Compounds

[0066] Jasmonic acid compounds employed in the invention includejasmonic acid and jasmonic acid derivatives available to one of skill inthe art. Such compounds include jasmonic acid, methyl jasmonate andtheir isomers. In the present invention jasmonic acid and jasmonic acidderivatives used also include synthetic and natural stereoisomers ofJasmonic acid, dihydrojasmonic acid, hydroxy jasmonic acid anddihydro-hydroxy jasmonic acid. Further examples of jasmonic acidderivatives that may be used in the invention include compounds havingany one of formulae VI.

[0067] wherein:

[0068] R3 is alkyl;

[0069] R4 is COOR, or —(CH₂)n-OX, where n is an integer of from 1 to 20;

[0070] R is H, or alkyl and

[0071] X is H, or 1 to 6 sugar residues (e.g., hexose or pentose).

[0072] In general, the alkyl groups employed in these jasmonic acidcompounds have about one to twenty carbon atoms, although in someembodiments lower alkyl groups are used, for example, alkyl groups withabout one to eight carbon atoms. Alkyl groups with even lower numbers ofcarbon atoms can also be used, for example, alkyl groups with one tosix, or one to three carbon atoms.

[0073] In some embodiments, jasmonic acid is employed in thecompositions of the invention. Jasmonic acid is a compound of formula VIwherein R3 is C₂H₅ and R4 is COOH.

[0074] Another jasmonic acid compound employed in the invention is acompound of formula VII.

[0075] wherein:

[0076] R3 is alkyl;

[0077] R4 is COOR, or —(CH₂)n-OX, where n is an integer of from 1 to 20;

[0078] R is H, or alkyl; and

[0079] X is H, or 1 to 6 sugar residues (e.g., hexoses or pentoses).

[0080] In some embodiments, dihydrojasmonic acid is employed in thecompositions of the invention. Dihydrojasmonic acid is a compound offormula VII wherein R3 is C₂H₅ and R4 is COOH.

[0081] Another jasmonic acid compound employed in the invention is acompound of formula VIII.

[0082] wherein:

[0083] R3 is alkyl;

[0084] R4 is COOR, or —(CH₂)n-OX, where n is an integer of from 1 to 20;

[0085] R is H, or alkyl;

[0086] X is H, or 1 to 6 sugar residues (e.g., hexoses or pentoses); and

[0087] Y is H, alkyl, or 1 to 6 sugar residues (e.g., hexoses orpentoses).

[0088] In some embodiments, hydroxyjasmonic acid is employed in thecompositions of the invention. Hydroxyjasmonic acid is a compound offormula VIII wherein R3 is C₂H₅ and R4 is COOH.

[0089] Another jasmonic acid compound employed in the invention is acompound of formula IX.

[0090] wherein:

[0091] R3 is alkyl;

[0092] R4 is COOR, or —(CH₂)n-OX, where n is an integer of from 1 to 20;

[0093] R is H, or alkyl;

[0094] X is H, or 1 to 6 sugar residues (e.g., hexoses or pentoses); and

[0095] Y is H, alkyl, or 1 to 6 sugar residues (e.g., hexoses orpentoses).

[0096] In some embodiments, dihydro-hydroxyjasmonic acid is employed inthe compositions of the invention. Dihydro-hydroxyjasmonic acid is acompound of formula IX wherein R3 is C₂H₅ and R4 is COOH.

[0097] When present in the compositions of the present invention,jasmonic acids or jasmonic acid derivatives can be used in an amount offrom about 0.001% to about 50%, or from about 0.01% to about 20%, orfrom about 0.01% to about 10%, or from about 0.05% to about 5%, or fromabout 0.05% to about 2% of the composition. According to the invention,in situ concentrations of jasmonic acid ranging from about 10⁻⁴ M toabout 10⁻⁶ M are effective for increasing cell proliferation andstimulating the production of collagen in mammalian keratinous tissue.

[0098] Zeatin Compounds

[0099] Zeatin compounds employed in the invention include the cis andtrans isomers of zeatin and the cis and trans isomers of zeatinderivatives available to one of skill in the art. Such zeatin compoundsand zeatin derivatives can be natural and synthetic derivatives of thecompounds provided by formula X below.

[0100] wherein:

[0101] R5 is H, 3-hydroxymethyl-3-methylallyl, alkyl, —(CH₂)n-CH₃, orOZ;

[0102] Z is H, 1 to 6 sugar residues (e.g., hexoses or pentoses), or—(CH₂)n-furan;

[0103] R6 is H, 3-hydroxymethyl-3-methylallyl, alkyl, —(CH₂)n-CH₃, orOZ; and

[0104] n is an integer of from about 1 to about 20.

[0105] In some embodiments, zeatin is employed in the compositions ofthe invention. Zeatin is a compound of formula X wherein R5 is3-hydroxymethyl-3-methylallyl, and R6 is H.

[0106] When present in the compositions of the present invention, zeatinor its derivatives can be used in an amount of from about 0.001% toabout 50%, or from about 0.01% to about 20%, or from about 0.01% toabout 10%, or from about 0.05% to about 5%, or from about 0.05% to about2% of the composition. According to the invention, in situconcentrations of zeatin ranging from about 10⁻⁴ M to about 10⁻⁶ M areeffective for increasing cell proliferation and stimulating theproduction of collagen in mammalian keratinous tissue.

[0107] Effects of the Compositions on Keratinous Tissues

[0108] The compositions and methods of the invention are useful forstimulating cellular proliferation and/or collagen production in cellswithin keratinous tissues. Such proliferation and collagen productioncan therapeutically improve visible and/or tactile discontinuities inmammalian skin, including discontinuities in skin texture and color. Forexample, after exposure to the compositions of the invention, theapparent diameter of pores can decrease, the apparent height of tissueimmediately proximate to pore openings approaches that of theinteradnexal skin, the skin tone/color becomes more uniform, and/or thelength, depth, and/or other dimension of lines and/or wrinkles aredecreased.

[0109] The compositions of the present invention are also useful forregulating the condition of skin and especially for regulatingkeratinous tissue condition. Regulation of skin condition, namelymammalian and in particular human skin condition, is often required dueto conditions that may be induced or caused by factors internal and/orexternal to the body. Examples include, environmental damage, radiationexposure (including ultraviolet radiation), chronological aging,menopausal status (e.g., post-menopausal changes in skin), stress,diseases, etc. For instance, “regulating skin condition” includesprophylactically regulating and/or therapeutically regulating skincondition, and may involve one or more of the following benefits:thickening of skin (i.e., building the epidermis, dermis, sub-dermal,subcutaneous fat, or underlying muscle layers of the skin) and, whereapplicable the keratinous layers of the nail and hair shaft, to reduceskin atrophy. The compositions and methods of the invention can alsoincrease the convolution of the dermal-epidermal border (also known asthe rete ridges), prevent loss of skin elasticity resulting from loss,damage and/or inactivation of functional skin elastin, prevent such aselastosis, sagging, loss of skin recoil from deformation; non-melaninskin discoloration such as under eye circles, blotching (e.g., unevenred coloration due to, e.g., rosacea) (hereinafter referred to as “redblotchiness”), sallowness (pale color), discoloration caused bytelangiectasia or spider vessels.

[0110] As used herein, prophylactically regulating skin conditionincludes delaying, minimizing and/or preventing visible and/or tactilediscontinuities in skin (e.g., texture irregularities in the skin whichmay be detected visually or by feel).

[0111] As used herein, therapeutically regulating skin conditionincludes ameliorating, e.g., diminishing, minimizing and/or effacing,discontinuities in skin.

[0112] Dental Applications

[0113] The invention also contemplates compositions for stimulatingcellular proliferation of gum tissues. Such compositions andformulations are useful for treating or preventing a variety of dentaland oral conditions, including gum recession, gingivitis and gumdisease. These compositions can contain an effective amount of a hydroxyacid compound, a jasmonic acid compound, a gibberellic acid compound ora zeatin compound as well as an orally acceptable carrier and otheringredients. Such compositions can be formulated as mouthwashes, as agel for use in a dental tray that can cup the teeth or as a adhesivethat can stick to tooth or gum surfaces.

[0114] In some embodiments, the carrier will include a tackifying agentand a solvent, which together yield a sticky matrix material. The matrixmaterial can be sufficiently sticky to enable a dental tray to be heldand retained against a person's teeth. Suitable sticky matrix materialsare preferably viscous and do not readily dissolve in saliva. Varioustackifying agents are available and the selection of the tackifyingagent can readily be made by one of skill in the art. One tackifyingagent that can be used to form a sticky and viscous matrix materialcomprises carboxypolymethylene, for example, CARBOPOL 934P.Carboxypolymethylene can be used to form a glue-like dental compositionthat itself can act as an adhesive in holding a comfortable,non-self-retaining dental tray against a person's teeth. The use ofcarboxypolymethylene eliminates the need to use dental trays that areself-retaining (i e., typically trays that are rigid and whichmechanically interlock over a person's teeth or gums and which areintended for use with less sticky compositions). See U.S. Pat. No.6,309,625.

[0115] In general, the dental compositions of the present invention caninclude carboxy-polymethylene in a concentration in a range from about0.5% to about 25% by weight of the dental composition, or in a rangefrom about 2% to about 12% and or in a range from about 3% to about 10%.Where is it desired to increase the stickiness, viscosity and resistanceto dilution to saliva, one may adjust the concentration ofcarboxypolymethylene to achieve a desired level of any or all of theseproperties. Increased stickiness assists in retaining the preferreddental trays against a person's teeth. Alternatively, compositions canbe made less adhesive and tacky if desired, particularly is applieddirectly without a dental tray.

[0116] In order to obtain good dispersion of the carboxypolymethyleneresin within the dental composition, it is recommended that thecarboxypolymethylene be mixed with a suitable solvent before attemptingto add other components that are less compatible withcarboxypolymethylene, such as water. Examples of suitable solvents foruse with carboxypolymethylene include glycerin, other polyhydricalcohols, polyalkylene glycols and other polyols, and the like. Glycerinappears to enable larger quantities of carboxypolymethylene to bedispersed in water. It is preferable that the concentration of glycerin,polyol, or like substance utilized as a solvent in the dental whiteningcompositions be added in a range from about 15% to about 85% by weightof the dental whitening compositions, more preferably in a range fromabout 25% to about 75% by weight, and most preferably in a range fromabout 30% to about 65% by weight. It should be understood, however, thatthe actual amount of carboxypolymethylene is not critical for obtaininga sticky, viscous dental composition.

[0117] The sticky matrix material may include other tackifyingcomponents that in combination with, or in lieu of some or all of, thecarboxypolymethylene will yield a gum stimulating composition having thedesired level of stickiness needed to hold a preferred,comfortable-fitting dental tray in place over a person's teeth. Othersynthetic polymers and/or natural gums, proteins, or other gel-formingadmixtures can be used so long as they yield a sticky gum stimulatingcomposition.

[0118] In addition to carboxypolymethylene, examples of other suitabletackifying and thickening agents include gums such as xanthan gum, talhagum, tragacanth gum, locust bean gum, guar gum, Irish moss gum, ghattigum, furcelleran gum, carrageenan gum, arabic gum, alginic acid gum,agar gum, alginate gum. Another suitable tackifying agent is sold asPEMULEN™, a proprietary compound from B. F. Goodrich, or a compositionalor chemical equivalent thereof. PEMULEN™ includes a significant quantityof a polyacrylic copolymer that has a slightly hydrophobic end and astrongly hydrophilic end. Additional examples of suitable tackifyingagents include polyethylene oxides such as POLYOX™ sold by UnionCarbide. These tackifying agents may be present in the same ranges asdiscussed above in relation to carboxypolymethylene.

[0119] One of skill in the art may include other active dental agents totreat or prevent other types of dental and/or gum problems. For example,in conjunction with the gum-stimulating components of the invention,such a skilled artisan may provide anti-cariogenic andanti-demineralizing agents such as fluoride salts, more particularlysodium monofluorophosphate, sodium fluoride, and stannous fluoride.Depending on the level of fluoride treatment desired, and depending onwhether or not a composition is “over-the-counter” or “by prescription”,the fluoride will be included in a range from about 0% to about 1% byweight of the dental whitening composition, more preferably in a rangefrom about 0.1% to about 0.5% by weight. Antimicrobial agents, e.g., forfighting gum disease, may be included in conjunction with the gumstimulating components of the invention. Examples of usefulantimicrobial agents include chlorohexidine, tetracycline, cetylpyridinium chloride, benzalkonium chloride, cetyl pyridinium bromide,methyl benzoate, and propyl benzoate. The antimicrobial agents arepreferably included in an amount in a range from about 0% to about 15%with the gum stimulating, or in a range from about 1% to about 5% byweight.

[0120] One method of dispensing sticky and viscous gum stimulatingcompositions within the scope of the present invention is by means of asyringe. Squeezable tubes and other similar dispensing devices may alsobe used to dispense the compositions. Upon dispensing, the gumstimulating compositions are sufficiently viscous that they do noteasily settle or spread once dispensed, but will generally remain as asingle extruded strand or bead of gum stimulating composition, forexample, along the gum line. Moreover, bottles, tubes or otherdispensing means known in the art may be used, particularly where thegum stimulating composition has lower viscosity, low stickiness, anddoes not include a thickening agent.

[0121] In some embodiments, the invention provides a unit dose of thegum stimulating compositions in a syringe or similar dispensing device.In this way, the person can load the precise amount of gum stimulatingcomposition onto the dental tray for each treatment period. By usingsuch dispensing devices, the dentist is also able to monitor how manydoses the person has received and used. In other embodiments, however,the gum stimulating compositions can be applied directly to the person'steeth without a dental tray, or a less viscous and sticky stimulatingcomposition according to the invention may be used in conjunction withself-retaining trays known in the art.

[0122] While a given gum stimulating composition may be able to retainthe dental tray against a person's teeth for, e.g., 10 hours or more,that composition could certainly be used within the scope of the presentinvention for any desired time period, such as for 15 minutes, one hour,or any desired time duration. In order to maximize treatment time andreduce the inconvenience of having a dental tray lodged within aperson's mouth the dental trays can be used at night during a person'ssleep.

[0123] Additional Ingredients

[0124] The compositions of the present invention may contain one or moreadditional skin care or gum care active ingredients.

[0125] In general, the additional components should be suitable forapplication to keratinous tissue, particularly when the composition isto be in contact with human keratinous tissue. Hence, the additionalingredients incorporated into the composition are suitable for contactwith human keratinous tissue and do out have undue toxicity,incompatibility, instability, allergic response, and the like within thescope of sound medical judgment.

[0126] The CTFA Cosmetic Ingredient Handbook, Second Edition (1992)describes a wide variety of nonlimiting cosmetic and pharmaceuticalingredients commonly used in the skin care industry, which are suitablefor use in the compositions of the present invention. Examples of theseingredient classes include: abrasives, absorbents, aesthetic componentssuch as fragrances, pigments, colorings/colorants, essential oils, skinsensates, astringents (clove oil, menthol, camphor, eucalyptus oil,eugenol, menthyl lactate, witch hazel distillate), anti-acne agents,anti-caking agents, antifoaming agents, antimicrobial agents,antioxidants, binders, biological additives, buffering agents, bulkingagents, chelating agents, chemical additives, colorants, cosmeticastringents, cosmetic biocides, denaturants, drug astringents, externalanalgesics, film formers, opacifying agents, pH adjusters, propellants;reducing agents, sequestrants, skin bleaching and lightening agents(e.g., hydroquinone, kojic acid, ascorbic acid, magnesium ascorbylphosphate, ascorbyl glucosamine), skin-conditioning agents (e.g.,humectants, including miscellaneous and occlusive), skin soothing and/orhealing agents (e.g., panthenol and panthenol derivatives), aloe vera,pantothenic acid, pantothenic acid derivatives, allantoin, bisabolol,and dipotassium glycyrrhizinate, skin treating agents, thickeners, andvitamins and derivatives thereof.

[0127] In any embodiment of the present invention, however, the activeingredients useful herein can be categorized by the benefit they provideor by their postulated mode of action. However, it is to be understoodthat the active ingredients useful herein can in some instances providemore than one benefit or operate via more than one mode of action.Therefore, classifications herein are made for the sake of convenienceand are not intended to limit the active to that particular applicationor applications listed.

[0128] Desquamation Compounds

[0129] A safe and effective amount of a desquamation compound may beadded to the compositions of the present invention, or from about 0.1%to about 10%, or from about 0.2% to about 5%, or from about 0.5% toabout 4%, by weight of the composition. Desquamation compounds enhancethe skin appearance benefits of the present invention. For example, thedesquamation compounds tend to improve the texture of the skin (e.g.,smoothness). One desquamation system that is suitable for use hereincontains sulfhydryl compounds and zwitterionic surfactants and isdescribed in U.S. Pat. No. 5,681,852, to Bissett, incorporated herein byreference. Zwitterionic surfactants such as described in theseapplications are also useful as desquamatory agents herein, with cetylbetaine being particularly desirable.

[0130] Anti-Acne Ingredients

[0131] The compositions of the present invention may contain a safe andeffective amount of one or more anti-acne active ingredients. Examplesof useful anti-acne ingredients include resorcinol, sulfur, benzoylperoxide, erythromycin, zinc, etc. Further examples of suitableanti-acne compounds are described in further detail in U.S. Pat. No.5,607,980, issued to McAtee et al, on Mar. 4, 1997.

[0132] Anti-Wrinkle Compounds/Anti-Atrophy Compounds

[0133] The compositions of the present invention may further contain asafe and effective amount of one or more anti-wrinkle compounds oranti-atrophy compounds. Exemplary anti-wrinkle/anti-atrophy compoundssuitable for use in the compositions of the present invention includesulfur-containing D and L amino acids and their derivatives and salts,particularly the N-acetyl derivatives, a preferred example of which isN-acetyl-L-cysteine; thiols (e.g. ethane thiol); phytic acid, lipoicacid; lysophosphatidic acid, skin peel agents (e.g., phenol and thelike), vitamin B₃ compounds and retinoids that enhance the health and/orappearance of keratinous tissues.

[0134] Vitamin B₃ Compounds

[0135] The compositions of the present invention may contain a safe andeffective amount of a vitamin B₃ compound. When vitamin B₃ compounds arepresent in the compositions of the instant invention, the compositionscan contain from about 0.01% to about 50%, or from about 0.1% to about10%, or from about 0.5% to about 10%, or from about 1% to about 5%, orfrom about 2% to about 5%, by weight of the composition, of the vitaminB₃ compound.

[0136] As used herein, “vitamin B₃ compound” means a compound having theformula:

[0137] wherein R is —CONH₂ (e.g., niacinamide), —COOH (e.g., nicotinicacid) or —CH₂OH (e.g., nicotinyl alcohol), derivatives thereof, andsalts of any of the foregoing. Exemplary derivatives of the foregoingvitamin B₃ compounds include nicotinic acid esters, includingnon-vasodilating esters of nicotinic acid (e.g., tocopheryl nicotinate),nicotinyl amino acids, nicotinyl alcohol esters of carboxylic acids,nicotinic acid N-oxide and niacinamide N-oxide.

[0138] The vitamin compounds may be included as the substantially purematerial, or as an extract obtained by suitable physical and/or chemicalisolation from natural (e.g., plant) sources.

[0139] Examples of suitable vitamin B₃ compounds are well known in theart and are commercially available from a number of sources, e.g., theSigma Chemical Company (St. Louis, Mo.); ICN Biomedicals, Inc. (Irvin,Calif.) and Aldrich Chemical Company (Milwaukee, Wis.).

[0140] Retinoids

[0141] The compositions of the present invention may also contain aretinoid. As used herein, “retinoid” includes all natural and/orsynthetic analogs of Vitamin A or retinol-like compounds that possessthe biological activity of Vitamin A in the skin as well as thegeometric isomers and stereoisomers of these compounds. The retinoidcan, for example, be retinol, retinol esters (e.g., C₂-C₂₂ alkyl estersof retinol, including retinyl palmitate, retinyl acetate, retinylpropionate), retinal, and/or retinoic acid (including all-trans retinoicacid and/or 13-cis-retinoic acid). In some embodiments, retinoids otherthan retinoic acid are used. These compounds are available in the artand are commercially available from a number of sources, e.g., SigmaChemical Company (St. Louis, Mo.), and Boerhinger Mannheim(Indianapolis, Ind.). Other retinoids that are useful herein aredescribed in U.S. Pat. No. 4,677,120, issued Jun. 30, 1987 to Parish etal.; U.S. Pat. No. 4,885,311, issued Dec. 5, 1989 to Parish et al.; U.S.Pat. No. 5,049,584, issued Sep. 17, 1991 to Purcell et al.; U.S. Pat.No. 5,124,356, issued Jun. 23, 1992 to Purcell et al.; and U.S. Pat. No.Reissue 34,075, issued Sep. 22, 1992 to Purcell et al. Other suitableretinoids are tocopheryl-retinoate, tocopherol ester of cis- ortrans-retinoic acid, adapalene(6-[3-(1-adamantyl)-4-methoxyphenyl]-2-naphthoic acid), and tazarotene(ethyl 6-[2-(4,4-dimethylthiochroman-6-yl)-ethynyl]nicotinate).Desirable retinoids include retinol, retinyl palmitate, retinyl acetate,retinyl propionate, retinal and combinations thereof.

[0142] The retinoid may be included as the substantially pure material,or as an extract obtained by suitable physical and/or chemical isolationfrom natural (e.g., plant) sources. In some embodiments, the retinoid issubstantially pure, or essentially pure.

[0143] The compositions of this invention may contain a safe andeffective amount of the retinoid, such that the resultant composition issafe and effective for regulating or improving the condition ofkeratinous tissues. The compositions and methods of the invention canimprove visible and/or tactile discontinuities in skin, or improve signsof skin aging. The compositions preferably contain from about 0.005% toabout 2%, or from about 0.01% to about 2%, retinoid. Retinol can also beused in an amount of from about 0.01% to about 0.15%. Retinol esters canbe used in an amount of from or about 0.01% to or about 2% (e.g., about1%). Retinoic acids can be used in an amount of from or about 0.01% toor about 0.25%. Tocopheryl-retinoate, adapalene, and tazarotene can beused in an amount of from or about 0.01% to or about 2%.

[0144] Where the compositions of the present invention contain both aretinoid and a vitamin B₃ compound, the retinoid can be used in theabove amounts, and the vitamin B₃ compound can be used in an amount offrom or about 0.1% to about 10%, or from about 2% to about 5%.

[0145] Peptides

[0146] Peptides, including but not limited to, di-, tri-, tetra-, andpentapeptides and derivatives thereof, may be included in thecompositions of the present invention in amounts that are safe andeffective. As used herein, “peptides” refers to both the naturallyoccurring peptides and synthesized peptides. Also useful herein arenaturally occurring and commercially available compositions that containpeptides.

[0147] Suitable dipeptides for use herein include Carnosine(beta-ala-his). Suitable tripeptides for use herein include,gly-his-lys, arg-lys-arg, and his-gly-gly. Preferred tripeptides andderivatives thereof include palmitoyl-gly-his-lys, which may bepurchased as Biopeptide CL™ (100 ppm of palmitoyl-gly-his-lyscommercially available from Sederma, France); Peptide CK (arg-lys-arg);Peptide CK+ (ac-arg-lys-arg-NH₂); and a copper derivative of his-gly-glysold commercially as lamin, from Sigma (St. Louis, Mo.). Suitabletetrapeptides for use herein include Peptide E, arg-ser-arg-lys (SEQ IDNO:1). Suitable pentapeptides for use herein include lys-thr-thr-lys-ser(SEQ ID NO:2). A preferred commercially available pentapeptidederivative composition is Matrixyl™, which contains 100 ppmpalmitoyl-lys-thr-thr-lys-ser (SEQ ID NO:3, commercially available fromSederma France).

[0148] Desirable peptides include palmitoyl-lys-thr-thr-lys-ser,palmitoyl-gly-his-lys, beta-ala-his, their derivatives, and combinationsthereof. In some embodiments, the peptide is selected frompalmitoyl-lys-thr-thr-lys-ser (SEQ ID NO:3), palmitoyl-gly-his-lys (SEQID NO:4), their derivatives, and combinations thereof. In otherembodiments, the peptide is selected from palmitoyl-lys-thr-thr-lys-ser(SEQ ID NO:3) and derivatives thereof.

[0149] When included in the present compositions, peptides can bepresent in amounts of from about 1×10⁻⁶% to about 10%, or from about1×10⁻⁶% to about 0.1%, or from about 1×10⁻⁵% to about 0.01%, by weightof the composition. In certain compositions where the peptide isCarnosine™, the compositions can contain from about 0.1% to about 5%, byweight of the composition, of such peptides. In other embodimentswherein the peptide-containing compositions, Matrixyl™, and/orBiopeptide CL™ are included, the compositions can contain from about0.1% to about 10%, by weight compositions, of Matrixyl™ and/orBiopeptide CL™ peptide-containing compositions.

[0150] Anti-Oxidants/Radical Scavengers

[0151] The compositions of the present invention may include a safe andeffective amount of an anti-oxidant/radical scavenger. Theanti-oxidant/radical scavenger is especially useful for providingprotection against ultraviolet radiation that can cause increasedscaling or texture changes in the stratum corneum and against otherenvironmental agents that can cause skin damage.

[0152] A safe and effective amount of an anti-oxidant/radical scavengermay be added to the compositions of the subject invention, for example,from about 0.1% to about 10%, or from about 1% to about 5%, of thecomposition.

[0153] Anti-oxidants/radical scavengers such as ascorbic acid (vitaminC) and its salts, ascorbyl esters of fatty acids, ascorbic acidderivatives (e.g., magnesium ascorbyl phosphate, sodium ascorbylphosphate, ascorbyl sorbate), tocopherol (vitamin E), tocopherolsorbate, tocopherol acetate, other esters of tocopherol, butylatedhydroxy benzoic acids and their salts,6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (commerciallyavailable under the trade name Trolox™), gallic acid and its alkylesters, especially propyl gallate, uric acid and its salts and alkylesters, sorbic acid and its salts, lipoic acid, amines (e.g.,N,N-diethylhydroxylamine, amino-guanidine), sulfhydryl compounds (e.g.,glutathione), dihydroxy fumaric acid and its salts, lysine pidolate,arginine pilolate, nordihydroguaiaretic acid, bioflavonoids, curcumin,lysine, methionine, proline, superoxide dismutase, silymarin, teaextracts, grape skin/seed extracts, melanin, and rosemary extracts maybe used. Other anti-oxidants/radical scavengers are selected fromtocopherol sorbate and other esters of tocopherol. For example, the useof tocopherol sorbate in topical compositions and applicable to thepresent invention is described in U.S. Pat. No. 4,847,071.

[0154] Chelators

[0155] The compositions of the present invention may also contain a safeand effective amount of a chelator or chelating agent. As used herein,“chelator” or “chelating agent” means an active agent capable ofremoving a metal ion from a system by forming a complex so that themetal ion cannot readily participate in or catalyze chemical reactions.The inclusion of a chelating agent is useful for providing protectionagainst ultraviolet radiation that can contribute to excessive scalingor skin texture changes and against other environmental agents that cancause skin damage.

[0156] A safe and effective amount of a chelating agent may be added tothe compositions of the subject invention, for example, from about 0.1%to about 10%, or from about 1% to about 5%, of the composition.Exemplary chelators that are useful herein are disclosed in U.S. Pat.No. 5,487,884; International Publication No. 91/16035, Bush et al.,published Oct. 31, 1995; and International Publication No. 91/16034,Bush et al., published Oct. 31, 1995. In some embodiments, the chelatorsused in compositions of the subject invention include, for example,furildioxime, furilmonoxime, and derivatives thereof.

[0157] Flavonoids

[0158] The compositions of the present invention may optionally containa flavonoid compound. Flavonoids are broadly disclosed in U.S. Pat. Nos.5,686,082 and 5,686,367, both of which are herein incorporated byreference. Flavonoids suitable for use in the present invention areflavanones selected from unsubstituted flavanones, mono-substitutedflavanones, and mixtures thereof; chalcones selected from unsubstitutedchalcones, mono-substituted chalcones, di-substituted chalcones,tri-substituted chalcones, and mixtures thereof; flavones selected fromunsubstituted flavones, mono-substituted flavones, di-substitutedflavones, and mixtures thereof; one or more isoflavones; coumarinsselected from unsubstituted coumarins, mono-substituted coumarins,di-substituted coumarins, and mixtures thereof; chromones selected fromunsubstituted chromones, mono-substituted chromones, di-substitutedchromones, and mixtures thereof; one or more dicoumarols; one or morechromanones; one or more chromanols; isomers (e.g., cis/trans isomers)thereof; and mixtures thereof. By the term “substituted” as used hereinmeans flavonoids wherein one or more hydrogen atom of the flavonoid hasbeen independently replaced with hydroxyl, C₁-C₈ alkyl, C₁-C₄ alkoxyl,O-glycoside, and the like or a mixture of these substituents.

[0159] Examples of suitable flavonoids include, but are not limited to,unsubstituted flavanone, mono-hydroxy flavanones (e.g., 2′-hydroxyflavanone, 6-hydroxy flavanone, 7-hydroxy flavanone, etc.), mono-alkoxyflavanones (e.g., 5-methoxy flavanone, 6-methoxy flavanone, 7-methoxyflavanone, 4′-methoxy flavanone, etc.), unsubstituted chalcone (e.g.unsubstituted trans-chalcone), mono-hydroxy chalcones (e.g., 2′-hydroxychalcone, 4′-hydroxy chalcone, etc.), di-hydroxy chalcones (e.g.,2′,4-dihydroxy chalcone, 2′,4′-dihydroxy chalcone, 2,2′-dihydroxychalcone, 2′,3-dihydroxy chalcone, 2′,5′-dihydroxy chalcone, etc.), andtri-hydroxy chalcones (e.g., 2′,3′,4′-trihydroxy chalcone,4,2′,4′-trihydroxy chalcone, 2,2′,4′-trihydroxy chalcone, etc.),unsubstituted flavone, 7,2′-dihydroxy flavone, 3′,4′-dihydroxynaphthoflavone, 4′-hydroxy flavone, 5,6-benzoflavone, and7,8-benzoflavone, unsubstituted isoflavone, daidzein (7,4′-dihydroxyisoflavone), 5,7-dihydroxy-4′-methoxy isoflavone, soy isoflavones (amixture extracted from soy), unsubstituted coumarin, 4-hydroxy coumarin,7-hydroxy coumarin, 6-hydroxy-4-methyl coumarin, unsubstituted chromone,3-formyl chromone, 3-formyl-6-isopropyl chromone, unsubstituteddicoumarol, unsubstituted chromanone, unsubstituted chromanol, andmixtures thereof.

[0160] In some embodiments, unsubstituted flavanone, methoxy flavanones,unsubstituted chalcone, 2′,4-dihydroxy chalcone, and mixtures thereofare used in the compositions of the invention. In other embodiments,unsubstituted flavanone, unsubstituted chalcone (especially the transisomer), and mixtures thereof are used in the compositions of theinvention.

[0161] Flavonoids can be synthesized or obtained as extracts fromnatural sources (e.g., plants). The naturally sourced material can alsofurther be derivatized (e.g., an ester or ether derivative preparedfollowing extraction from a natural source). Flavonoid compounds usefulherein are also commercially available from a number of sources, e.g.,Indofine Chemical Company, Inc. (Somerville, N.J.), Steraloids, Inc.(Wilton, N.H.), and Aldrich Chemical Company, Inc. (Milwaukee, Wis.).Mixtures of such flavonoid compounds may also be used.

[0162] The flavonoid compounds can be present in the invention, forexample, at concentrations of from about 0.01% to about 20%, morepreferably from about 0.1% to about 10%, and still more preferably fromabout 0.5% to about 5%.

[0163] Anti-Inflammatory Agents

[0164] A safe and effective amount of an anti-inflammatory agent may beadded to the compositions of the present invention, for example, fromabout 0.1% to about 10%, or from about 0.5% to about 5%, of thecomposition. The anti-inflammatory agent can enhance the appearance ofthe skin, for example, by contributing to a more uniform and acceptableskin tone or color. The exact amount of anti-inflammatory agent to beused in the compositions will depend on the particular anti-inflammatoryagent utilized since such agents vary widely in potency.

[0165] Steroidal anti-inflammatory agents, including but not limited to,corticosteroids such as hydrocortisone, hydroxyltriamcinolone,alpha-methyl dexamethasone, dexamethasone-phosphate, beclomethasonedipropionates, clobetasol valerate, desonide, desoxymethasone,desoxycorticosterone acetate, dexamethasone, dichlorisone, diflorasonediacetate, diflucortolone valerate, fluadrenolone, flucloroloneacetonide, fludrocortisone, flumethasone pivalate, fluosinoloneacetonide, fluocinonide, flucortine butylesters, fluocortolone,fluprednidene (fluprednylidene) acetate, flurandrenolone, halcinonide,hydrocortisone acetate, hydrocortisone butyrate, methylprednisolone,triamcinolone acetonide, cortisone, cortodoxone, flucetonide,fludrocortisone, difluorosone diacetate, fluradrenolone,fludrocortisone, diflurosone diacetate, fluradrenolone acetonide,medrysone, amcinafel, amcinafide, betamethasone and the balance of itsesters, chloroprednisone, chlorprednisone acetate, clocortelone,clescinolone, dichlorisone, diflurprednate, flucloronide, flunisolide,fluoromethalone, fluperolone, fluprednisolone, hydrocortisone valerate,hydrocortisone cyclopentylpropionate, hydrocortamate, meprednisone,paramethasone, prednisolone, prednisone, beclomethasone dipropionate,triamcinolone, and mixtures thereof may be used. In some embodiments,the steroidal anti-inflammatory used is hydrocortisone.

[0166] A second class of anti-inflammatory agents that is useful in thecompositions includes the nonsteroidal anti-inflammatory agents. Avariety of compounds are encompassed by this group. For detaileddisclosure of the chemical structure, synthesis, side effects, etc. ofnon-steroidal anti-inflammatory agents, one may refer to standard texts,including Anti-inflammatory and Anti-Rheumatic Drugs, K. D. Rainsford,Vol. I-III, CRC Press, Boca Raton, (1985), and Anti-inflammatory Agents,Chemistry and Pharmacology, 1, R. A. Scherrer, et al., Academic Press,New York (1974).

[0167] Specific non-steroidal anti-inflammatory agents useful in thecomposition invention include, but are not limited to:

[0168] 1) the oxicams, such as piroxicam, isoxicam, tenoxicam,sudoxicam, and CP-14,304;

[0169] 2) benorylate, trilisate, safapryn, solprin, diflunisal, andfendosal;

[0170] 3) the acetic acid derivatives, such as diclofenac, fenclofenac,indomethacin, sulindac, tolmetin, isoxepac, furofenac, tiopinac,zidometacin, acematacin, fentiazac, zomepirac, clindanac, oxepinac,felbinac, and ketorolac;

[0171] 4) the fenamates, such as mefenamic, meclofenamic, flufenamic,niflumic, and tolfenamic acids;

[0172] 5) the propionic acid derivatives, such as ibuprofen, naproxen,benoxaprofen, flurbiprofen, ketoprofen, fenoprofen, fenbufen,indopropfen, pirprofen, carprofen, oxaprozin, pranoprofen, miroprofen,tioxaprofen, suprofen, alminoprofen, and tiaprofenic; and

[0173] 6) the pyrazoles, such as phenylbutazone, oxyphenbutazone,feprazone, azapropazone, and trimethazone.

[0174] Mixtures of these non-steroidal anti-inflammatory agents may alsobe employed, as well as the dermatologically acceptable salts and estersof these agents. For example, etofenamate, a flufenamic acid derivative,is useful for topical application. Of the nonsteroidal anti-inflammatoryagents, ibuprofen, naproxen, flufenamic acid, etofenamate, aspirin,mefenamic acid, meclofenamic acid, piroxicam and felbinac are oftenused. Ibuprofen, naproxen, ketoprofen, etofenamate, aspirin andflufenamic acid are frequently used.

[0175] Moreover, so-called “natural” anti-inflammatory agents are usefulin methods of the present invention. Such agents may suitably beobtained as an extract by suitable physical and/or chemical isolationfrom natural sources (e.g., plants, fungi, by-products ofmicroorganisms) or can be synthetically prepared. For example,candelilla wax, bisabolol (e.g., alpha bisabolol), aloe vera, plantsterols (e.g., phytosterol), Manjistha (extracted from plants in thegenus Rubia, particularly Rubia Cordifolia), and Guggal (extracted fromplants in the genus Commiphora, particularly Commiphora Mukul), kolaextract, chamomile, red clover extract, and sea whip extract, may beused.

[0176] Additional anti-inflammatory agents useful herein includecompounds of the Licorice (the plant genus/species Glycyrrhiza glabra)family, including glycyrrhetic acid, glycyrrhizic acid, and derivativesthereof (e.g., salts and esters). Suitable salts of the foregoingcompounds include metal and ammonium salts. Suitable esters includeC₂-C₂₄ saturated or unsaturated esters of the acids, or C₁₀-C₂₄, orC₁₆-C₂₄. Specific examples of the foregoing include oil soluble licoriceextract, the glycyrrhizic and glycyrrhetic acids themselves,monoammonium glycyrrhizinate, monopotassium glycyrrhizinate, dipotassiumglycyrrhizinate, 1-beta-glycyrrhetic acid, stearyl glycyrrhetinate, and3-stearyloxy-glycyrrhetinic acid, and disodium3-succinyloxy-beta-glycyrrhetinate. Stearyl glycyrrhetinate ispreferred.

[0177] Anti-Cellulite Agents

[0178] The compositions of the present invention may also contain a safeand effective amount of an anti-cellulite agent. Suitable agents mayinclude, but are not limited to, xanthine compounds (e.g., caffeine,theophylline, theobromine, and aminophylline).

[0179] Topical Anesthetics

[0180] The compositions of the present invention may also contain a safeand effective amount of a topical anesthetic. Examples of topicalanesthetic drugs include benzocaine, lidocaine, bupivacaine,chlorprocaine, dibucaine, etidocaine, mepivacaine, tetracaine,dyclonine, hexylcaine, procaine, cocaine, ketamine, pramoxine, phenol,and pharmaceutically acceptable salts thereof.

[0181] Tanning Compounds

[0182] The compositions of the present invention may contain a tanningcompound. When present, the compositions can contain from about 0.1% toabout 20%, or from about 2% to about 7%, or from about 3% to about 6%,by weight of the composition, of dihydroxyacetone as an artificialtanning compound.

[0183] Dihydroxyacetone, which is also known as DHA or1,3-dihydroxy-2-propanone, is a white to off-white, crystalline powder.This material can be represented by the chemical formula C₃H₆O₃. Thecompound can exist as a mixture of monomers and dimers, with the dimerspredominating in the solid crystalline state. Upon heating or melting,the dimers 25 break down to yield the monomers. This conversion of thedimeric form to the monomeric form also occurs in aqueous solution.Dihydroxyacetone is also known to be more stable at acidic pH values.See The Merck Index, Tenth Edition, entry 3167, p. 463 (1983), and“Dihydroxyacetone for Cosmetics”, E. Merck Technical Bulletin, 03-304110, 319 897, 180 588.

[0184] Skin Lightening Agents

[0185] The compositions of the present invention may contain askin-lightening agent. When used, the compositions can contain fromabout 0.1% to about 10%, or from about 0.2% to about 5%, or from about0.5% to about 2%, by weight of the composition, of a skin lighteningagent. Suitable skin lightening agents include those known in the art,including kojic acid, arbutin, ascorbic acid and derivatives thereof(e.g., magnesium ascorbyl phosphate or sodium ascorbyl phosphate), andextracts (e.g., mulberry extract, placental extract). Skin lighteningagents suitable for use herein also include those described in the PCTpublication No. 95/34280, in the name of Hillebrand, corresponding toPCT Application No. U.S. Ser. No. 95/07432, filed Jun. 12, 1995; andco-pending U.S. application Ser. No. 08/390,152 filed in the names ofKvalnes, Mitchell A. DeLong, Barton J. Bradbury, Curtis B. Motley, andJohn D. Carter, corresponding to PCT Publication No. 95/23780, publishedSep. 8, 1995.

[0186] Skin Soothing and Skin Healing Compounds

[0187] The compositions of the present invention may comprise a skinsoothing or skin-healing compound. Skin soothing or skin healingcompounds suitable for use herein include panthenoic acid derivatives(including panthenol, dexpanthenol, ethyl panthenol), aloe vera,allantoin, bisabolol, and dipotassium glycyrrhizinate. A safe andeffective amount of a skin soothing or skin healing compound may beadded to the present composition, foe example, from about 0.1% to about30%, or from about 0.5% to about 20%, or from about 0.5% to about 10%,by weight of the composition formed.

[0188] Anti-microbial and Anti-fungal Compounds

[0189] The compositions of the present invention may contain ananti-microbial or anti-fungal compound. Such compounds are capable ofdestroying microbes, preventing the development of microbes orpreventing the pathogenic action of microbes. A safe and effectiveamount of an anti-microbial or anti-fungal compound may be added to thepresent compositions, for example, from about 0.001% to about 10%, orfrom about 0.01% to about 5%, or from about 0.05% to about 2%.

[0190] Examples of antimicrobial and antifungal compounds includeβ-lactam drugs, quinolone drugs, ciprofloxacin, norfloxacin,tetracycline, erythromycin, amikacin, 2,4,4′-trichloro-2′-hydroxydiphenyl ether, 3,4,4′-trichlorobanilide, phenoxyethanol, phenoxypropanol, phenoxyisopropanol, doxycycline, capreomycin, chlorhexidine,chlortetracycline, oxytetracycline, clindamycin, ethambutol, hexamidineisethionate, metronidazole, pentamidine, gentamicin, kanamycin,lineomycin, methacycline, methenamine, minocycline, neomycin,netilmicin, paromomycin, streptomycin, tobramycin, miconazole,tetracycline hydrochloride, erythromycin, zinc erythromycin,erythromycin estolate, erythromycin stearate, amikacin sulfate,doxycycline hydrochloride, capreomycin sulfate, chlorhexidine gluconate,chlorhexidine hydrochloride, chlortetracycline hydrochloride,oxytetracycline hydrochloride, clindamycin hydrochloride, ethambutolhydrochloride, metronidazole hydrochloride, pentamidine hydrochloride,gentamicin sulfate, kanamycin sulfate, lineomycin hydrochloride,methacycline hydrochloride, methenamine hippurate, methenaminemandelate, minocycline hydrochloride, neomycin sulfate, netilmicinsulfate, paromomycin sulfate, streptomycin sulfate, tobramycin sulfate,miconazole hydrochloride, ketaconazole, amanfadine hydrochloride,amanfadine sulfate, octopirox, parachlorometa xylenol, nystatin,tolnaftate, zinc pyrithione and clotrimazole.

[0191] Examples of compounds useful herein include those selected frombenzoyl peroxide, 3-hydroxy benzoic acid, glycolic acid, lactic acid,4-hydroxy benzoic acid, 2-hydroxybutanoic acid, 2-hydroxypentanoic acid,2-hydroxyhexanoic acid, cis-retinoic acid, trans-retinoic acid, retinol,phytic acid, N-acetyl-L-cysteine, lipoic acid, azelaic acid, arachidonicacid, benzoylperoxide, tetracycline, ibuprofen, naproxen,hydrocortisone, acetominophen, resorcinol, phenoxyethanol,phenoxypropanol, phenoxyisopropanol, 2,4,4′-trichloro-2′-hydroxydiphenyl ether, 3,4,4′-trichlorocarbanilide, octopirox, lidocainehydrochloride, clotrimazole, miconazole, ketoconazole, neocycin sulfate,and mixtures thereof.

[0192] Sunscreen Compounds

[0193] Exposure to ultraviolet light can result in excessive scaling andtexture changes of the stratum corneum. Therefore, the compositions ofthe subject invention may optionally contain a sunscreen compound. Asused herein, “sunscreen compound” includes both sunscreen agents andphysical sun blocks. Suitable sunscreen compounds may be organic orinorganic.

[0194] Inorganic sunscreens useful herein include the following metallicoxides; titanium dioxide having an average primary particle size of fromabout 15 nm to about 100 nm, zinc oxide having an average primaryparticle size of from about 15 nm to about 150 nm, zirconium oxidehaving an average primary particle size of from about 15 nm to about 150nm, iron oxide having an average primary particle size of from about 15nm to about 500 nm, and mixtures thereof. When used herein, theinorganic sunscreens can be present in the amount of from about 0.1% toabout 20%, or from about 0.5% to about 10%, or from about 1% to about5%, by weight of the composition.

[0195] A wide variety of conventional organic sunscreen compounds aresuitable for use herein. Sagarin, et al., at Chapter VIII, pages 189 etseq., of Cosmetics Science and Technology (1972), discloses numeroussuitable compounds. Specific suitable sunscreen compounds include, forexample: p-aminobenzoic acid, its salts and its derivatives (ethyl,isobutyl, glyceryl esters; p-dimethylaminobenzoic acid); anthranilates(i.e., o-amino-benzoates; methyl, menthyl, phenyl, benzyl, phenylethyl,linalyl, terpinyl, and cyclohexenyl esters); salicylates esters (amyl,phenyl, octyl, benzyl, menthyl, glyceryl, and di-pro-pyleneglycolesters); cinnamic acid derivatives (menthyl and benzyl esters, a-phenylcinnamonitrile; butyl cinnamoyl pyruvate); dihydroxycinnamic acidderivatives (umbelliferone, methylumbelliferone,methylaceto-umbelliferone); trihydroxy-cinnamic acid derivatives(esculetin, methylesculetin, daphnetin, and the glucosides, esculin anddaphnin); hydrocarbons (diphenylbutadiene, stilbene); dibenzalacetoneand benzalacetophenone; naphtholsulfonates (sodium salts of2-naphthol-3,6-disulfonic and of 2-naphthol-6,8-disulfonic acids);di-hydroxynaphthoic acid and its salts; o- andp-hydroxybiphenyldisulfonates; coumarin derivatives (7-hydroxy,7-methyl, 3-phenyl); diazoles (2-acetyl-3-bromoindazole, phenylbenzoxazole, methyl naphthoxazole, various aryl benzothiazoles); quininesalts (bisulfate, sulfate, chloride, oleate, and tannate); quinolinederivatives (8-hydroxyquinoline salts, 2-phenylquinoline); hydroxy- ormethoxy-substituted benzophenones; uric and violuric acids; tannic acidand its derivatives (e.g., hexaethylether); (butyl carbotol) (6-propylpiperonyl) ether; hydroquinone; benzophenones (oxybenzene,sulisobenzone, dioxybenzone, benzoresorcinol,2,2,4,4′-tetrahydroxybenzophenone,2,2′-dihydroxy-4,4′-dimethoxybenzophenone, octabenzone;4-isopropyldibenzoylmethane; butylmethoxydibenzoylmethane; etocrylene;octocrylene; [3-(4′-methylbenzylidene bornan-2-one), terephthalylidenedicamphor sulfonic acid and 4-isopropyl-di-benzoylmethane.

[0196] Desirable compounds include 2-ethylhexyl-p-methoxycinnamate(commercially available as PARSOL MCX), 4,4′-t-butylmethoxydibenzoyl-methane (commercially available as PARSOL 1789),2-hydroxy-4-methoxybenzophenone, octyldimethyl-p-aminobenzoic acid,digalloyltrioleate, 2,2-dihydroxy-4-methoxybenzophenone,ethyl-4-(bis(hydroxy-propyl))aminobenzoate,2-ethylhexyl-2-cyano-3,3-diphenylacrylate, 2-ethylhexyl-salicylate,glyceryl-p-aminobenzoate, 3,3,5-tri-methylcyclohexylsalicylate,methylanthranilate, p-dimethyl-aminobenzoic acid or aminobenzoate,2-ethylhexyl-p-dimethyl-amino-benzoate, 2-phenylbenzimidazole-5-sulfonicacid, 2-(p-dimethylaminophenyl)-5-sulfonicbenzoxazoic acid, octocryleneand mixtures of these compounds.

[0197] In some embodiments, the organic sunscreen compounds used in thecompositions of the invention are 2-ethylhexyl-p-methoxycinnamate,butylmethoxydibenzoyl-methane, 2-hydroxy-4-methoxybenzo-phenone,2-phenylbenzimidazole-5-sulfonic acid, octyldimethyl-p-aminobenzoicacid, octocrylene and mixtures thereof.

[0198] Useful sunscreen compounds are also described in U.S. Pat. No.4,937,370 issued to Sabatelli on Jun. 26, 1990, and U.S. Pat. No.4,999,186 issued to Sabatelli & Spirnak on Mar. 12, 1991. Thesun-screening agents disclosed therein have, in a single molecule, twodistinct chromophore moieties that exhibit different ultra-violetradiation absorption spectra. One of the chromophore moieties absorbspredominantly in the UVB radiation range and the other absorbs stronglyin the UVA radiation range.

[0199] Desirable members of this class of sun-screening agents are4-N,N-(2-ethylhexyl)methyl-aminobenzoic acid ester of2,4-dihydroxybenzophenone; N,N-di-(2-ethylhexyl)-4-aminobenzoic acidester with 4-hydroxydibenzoylmethane;4-N,N-(2-ethylhexyl)methyl-aminobenzoic acid ester with4-hydroxydibenzoylmethane; 4-N,N-(2-ethylhexyl)methyl-aminobenzoic acidester of 2-hydroxy-4-(2-hydroxyethoxy)benzophenone;4-N,N-(2-ethylhexyl)-methylaminobenzoic acid ester of4-(2-hydroxyethoxy)dibenzoylmethane;N,N-di-(2-ethylhexyl)-4-aminobenzoic acid ester of2-hydroxy-4-(2-hydroxyethoxy)benzophenone; andN,N-di-(2-ethylhexyl)-4-aminobenzoic acid ester of4-(2-hydroxyethoxy)dibenzoylmethane and mixtures thereof. Otherdesirable sunscreen compounds include4,4′-t-butylmethoxydibenzoyl-methane, 2-ethylhexyl-p-methoxycinnamate,phenyl benzimidazole sulfonic acid, and octocrylene.

[0200] A safe and effective amount of the organic sunscreen compound isused, typically from about 1% to about 20%, more typically from about 2%to about 10% by weight of the composition. Exact amounts will varydepending upon the sunscreen or sunscreens chosen and the desired SunProtection Factor (SPF).

[0201] Particulate Material

[0202] The compositions of the invention may contain a particulatematerial, for example, a metallic oxide. These particulates can becoated or uncoated, charged or uncharged. Charged particulate materialsare disclosed in U.S. Pat. No. 5,997,887, to Ha, et al., incorporatedherein by reference. Particulate materials useful herein include;bismuth oxychloride, iron oxide, mica, mica treated with barium sulfateand TiO₂, silica, nylon, polyethylene, talc, styrene, polypropylene,ethylene/acrylic acid copolymer, sericite, titanium dioxide, bismuthoxychloride, iron oxide, aluminum oxide, silicone resin, barium sulfate,calcium carbonate, cellulose acetate, polymethyl methacrylate, andmixtures thereof.

[0203] Inorganic particulate materials, e.g., TiO₂, ZnO, or ZrO₂ arecommercially available from a number of sources. One example of asuitable particulate material contains the material available from U.S.Cosmetics (TRONOX TiO2 series, SAT-T CR837, a rutile TiO₂). Particulatematerials can be present in the composition in levels of from about0.01% to about 2%, or from about 0.05% to about 1.5%, or from about 0.1%to about 1%, by weight of the composition.

[0204] Conditioning Agents

[0205] The compositions of the present invention may contain aconditioning agent selected from humectants, moisturizers, or skinconditioners. A variety of these materials can be employed and each canbe present at a level of from about 0.01% to about 20%, more preferablyfrom about 0.1% to about 10%, and still more preferably from about 0.5%to about 7% by weight of the composition. These materials include, butare not limited to, guanidine; urea; glycolic acid and glycolate salts(e.g. ammonium and quaternary alkyl ammonium); lactic acid and lactatesalts (e.g., ammonium and quaternary alkyl ammonium); aloe vera in anyof its variety of forms (e.g., aloe vera gel); polyhydroxy alcohols suchas sorbitol, mannitol, xylitol, erythritol, glycerol, hexanetriol,butanetriol, propylene glycol, butylene glycol, hexylene glycol and thelike; polyethylene glycols; sugars (e.g., melibiose) and starches; sugarand starch derivatives (e.g., alkoxylated glucose, fucose, glucosamine);hyaluronic acid; lactamide monoethanolamine; acetamide monoethanolamine;panthenol; allantoin; and mixtures thereof. Also useful herein are thepropoxylated glycerols described in U.S. Pat. No. 4,976,953, to Orr etal, issued Dec. 11, 1990.

[0206] Also useful are various C₁-C₃₀ monoesters and polyesters ofsugars and related materials. These esters are derived from a sugar orpolyol moiety and one or more carboxylic acid moieties. Such estermaterials are further described in, U.S. Pat. No. 2,831,854; U.S. Pat.No. 4,005,196, to Jandacek, issued Jan. 25, 1977; U.S. Pat. No.4,005,195, to Jandacek, issued Jan. 25, 1977; U.S. Pat. No. 5,306,516,to Letton et al, issued Apr. 26, 1994; U.S. Pat. No. 5,306,515, toLetton et al, issued Apr. 26, 1994; U.S. Pat. No. 5,305,514, to Lettonet al, issued Apr. 26, 1994; U.S. Pat. No. 4,797,300, to Jandacek et al,issued Jan. 10, 1989; U.S. Pat. No. 3,963,699, to Rizzi et al, issuedJun. 15, 1976; U.S. Pat. No. 4,518,772, to Volpenhein, issued May 21,1985; and U.S. Pat. No. 4,517,360, to Volpenhein, issued May 21, 1985.

[0207] Desirable conditioning agents are selected from urea, guanidine,sucrose polyester, panthenol, dexpanthenol, allantoin, and combinationsthereof.

[0208] Thickening Agent

[0209] The compositions of the present invention can contain one or morethickening agents, can be from about 0.1% to about 5%, or from about0.1% to about 4%, or from about 0.25% to about 3%, by weight of thecomposition. Nonlimiting classes of thickening agents include thoseselected from the following:

[0210] a) Carboxylic Acid Polymers

[0211] These polymers are crosslinked compounds containing one or moremonomers derived from acrylic acid, substituted acrylic acids, and saltsand esters of these acrylic acids and the substituted acrylic acids,wherein the crosslinking agent contains two or more carbon-carbon doublebonds and is derived from a polyhydric alcohol. Polymers useful in thepresent invention are more fully described in U.S. Pat. No. 5,087,445,to Haffey et al, issued Feb. 11, 1992; U.S. Pat. No. 4,509,949, to Huanget al, issued Apr. 5, 1985; U.S. Pat. No. 2,798,053, to Brown, issuedJul. 2, 1957; and in CTFA International Cosmetic Ingredient Dictionary,Fourth Edition, 1991, pp. 12 and 80.

[0212] Examples of commercially available carboxylic acid polymersuseful herein include the carbomers, which are homopolymers of acrylicacid crosslinked with allyl ethers of sucrose or pentaerytritol. Thecarbomers are available as the Carbopol™ 900 series from B.F. Goodrich(e.g., Carbopol™ 954). In addition, other suitable carboxylic acidpolymeric agents include copolymers of C₁₀₋₃₀ alkyl acrylates with oneor more monomers of acrylic acid, methacrylic acid, or one of theirshort chain (i.e., C₁₋₄ alcohol) esters, wherein the crosslinking agentis an allyl ether of sucrose or pentaerytritol. These copolymers areknown as acrylates/C₁₀₋₃₀ alkyl acrylate crosspolymers and arecommercially available as Carbopol™ 1342, Carbopol™ 1382, Pemulen TR-1,and Pemulen TR-2, from B.F. Goodrich. In other words, examples ofcarboxylic acid polymer thickeners useful herein are those selected fromcarbomers, acrylates/C₁₀-C₃₀ alkyl acrylate crosspolymers, and mixturesthereof.

[0213] b) Crosslinked Polyacrylate Polymers

[0214] The compositions of the present invention can optionally containcrosslinked polyacrylate polymers useful as thickeners or gelling agentsincluding both cationic and nonionic polymers, with the cationics beinggenerally preferred. Examples of useful crosslinked nonionicpolyacrylate polymers and crosslinked cationic polyacrylate polymers arethose described in U.S. Pat. No. 5,100,660, to Hawe et al, issued Mar.31, 1992; U.S. Pat. No. 4,849,484, to Heard, issued Jul. 18, 1989; U.S.Pat. No. 4,835,206, to Farrar et al, issued May 30, 1989; U.S. Pat. No.4,628,078 to Glover et al issued Dec. 9, 1986; U.S. Pat. No. 4,599,379to Flesher et al issued Jul. 8, 1986; and EP 228,868, to Farrar et al,published Jul. 15, 1987.

[0215] c) Polyacrylamide Polymers

[0216] The compositions of the present invention can optionally containpolyacrylamide polymers, especially nonionic polyacrylamide polymersincluding substituted branched or unbranched polymers. More preferredamong these polyacrylamide polymers is the nonionic polymer given theCTFA designation polyacrylamide and isoparaffin and laureth-7, availableunder the Trade name Sepigel 305 from Seppic Corporation (Fairfield,N.J.).

[0217] Other polyacrylamide polymers useful herein include multi-blockcopolymers of acrylamides and substituted acrylamides with acrylic acidsand substituted acrylic acids. Commercially available examples of thesemulti-block copolymers include Hypan SR150H, SS500V, SS500W, SSSA100H,from Lipo Chemicals, Inc., (Patterson, N.J.).

[0218] d) Polysaccharides

[0219] A wide variety of polysaccharides are useful herein.“Polysaccharides” refer to gelling agents that contain a backbone ofrepeating sugar (i.e., carbohydrate) units. Nonlimiting examples ofpolysaccharide gelling agents include those selected from cellulose,carboxymethyl hydroxyethylcellulose, cellulose acetate propionatecarboxylate, hydroxyethylcellulose, hydroxyethyl ethylcellulose,hydroxypropylcellulose, hydroxypropyl methylcellulose, methylhydroxyethylcellulose, microcrystalline cellulose, sodium cellulosesulfate, and mixtures thereof. Also useful herein are thealkyl-substituted celluloses. In these polymers, the hydroxy groups ofthe cellulose polymer is hydroxyalkylated (preferably hydroxyethylatedor hydroxypropylated) to form a hydroxyalkylated cellulose that is thenfurther modified with a C₁₀-C₃₀ straight chain or branched chain alkylgroup through an ether linkage. Typically these polymers are ethers ofC₁₀-C₃₀ straight or branched chain alcohols with hydroxyalkylcelluloses.Examples of alkyl groups useful herein include those selected fromstearyl, isostearyl, lauryl, myristyl, cetyl, isocetyl, cocoyl (i.e.alkyl groups derived from the alcohols of coconut oil), palmityl, oleyl,linoleyl, linolenyl, ricinoleyl, behenyl, and mixtures thereof.Preferred among the alkyl hydroxyalkyl cellulose ethers is the materialgiven the CTFA designation cetyl hydroxyethylcellulose, which is theether of cetyl alcohol and hydroxyethylcellulose. This material is soldunder the trade name Natrosol™ CS Plus from Aqualon Corporation(Wilmington, Del.).

[0220] Other useful polysaccharides include scleroglucans that are alinear chain of (1-3) linked glucose units with a (1-6) linked glucoseevery three units, a commercially available example of which isClearogel™ CS11 from Michel Mercier Products Inc. (Mountainside, N.J.).

[0221] e) Gums

[0222] Other thickening and gelling agents useful herein includematerials that are primarily derived from natural sources. Nonlimitingexamples of these gelling agent gums include acacia, agar, algin,alginic acid, ammonium alginate, amylopectin, calcium alginate, calciumcarrageenan, carnitine, carrageenan, dextrin, gelatin, gellan gum, guargum, guar hydroxypropyltrimonium chloride, hectorite, hyaluroinic acid,hydrated silica, hydroxypropyl chitosan, hydroxypropyl guar, karaya gum,kelp, locust bean gum, natto gum, potassium alginate, potassiumcarrageenan, propylene glycol alginate, sclerotium gum, sodiumcarboyxmethyl dextran, sodium carrageenan, tragacanth gum, xanthan gum,and mixtures thereof.

[0223] Compositions of the invention can therefore include desirablethickening agents such as carboxylic acid polymers, crosslinkedpolyacrylate polymers, polyacrylamide polymers, and mixtures thereof,more preferably selected from carboxylic acid polymers, polyacrylamidepolymers, and mixtures thereof.

[0224] Composition Preparation

[0225] The compositions useful for the methods of the present inventionare generally prepared by conventional methods such as are known in theart of making topical compositions. Such methods typically involvemixing of the ingredients in one or more steps to a relatively uniformstate, with or without heating, cooling, application of vacuum, and thelike.

[0226] Methods for Regulating Skin Condition

[0227] The compositions of the present invention are useful forstimulating cellular growth and/or collagen production in keratinoustissues. Such increased cell growth and/or increased collagen productioncan help regulate or rejuvenate mammalian keratinous tissues. Thecompositions of the invention can be used for both prophylactic andtherapeutic treatment of skin conditions. For example, compositions ofthe invention can be used for wound healing, thickening keratinoustissue (i.e., building the epidermis and/or dermis layers of the skinand where applicable the keratinous layers of the nail and hair shaft),preventing and/or retarding atrophy of mammalian skin, preventing and/orretarding the appearance of spider vessels and/or red blotchiness onmammalian skin, preventing and/or retarding the appearance of darkcircles under the eye of a mammal, preventing and/or retardingsallow-colored mammalian skin, preventing and/or retarding sagging ofmammalian skin, softening and/or smoothing lips, hair and nails of amammal, preventing and/or relieving itch of mammalian skin, regulatingskin texture (e.g. wrinkles and fine lines), and improving skin color(e.g. redness, freckles).

[0228] Treating keratinous tissues involves topically applying to thekeratinous tissue a safe and effective amount of a composition of thepresent invention. The amount of the composition that is applied, thefrequency of application and the period of use will vary widelydepending upon the level of the acetylsalicylic acid, gibberellic acid,jasmonic acid, salicylic acid, or zeatin in a given composition and thelevel of regulation desired, for example, in light of the level ofkeratinous tissue damage present or expected to occur.

[0229] In some embodiments, the composition is chronically applied tothe skin. By “chronic topical application” is meant continued topicalapplication of the composition over an extended period during thesubject's lifetime, for example, for a period of at least about oneweek, or for a period of at least about one month, or for at least aboutthree months, or for at least about six months, or for at least aboutone year. While benefits are obtainable after various periods of use(e.g., five, ten or twenty years), chronic application can continuethroughout the subject's lifetime. Typically applications would be onthe order of about once per day over such extended periods, howeverapplication rates can vary from about once per week up to about threetimes per day or more.

[0230] A wide range of quantities of the compositions of the presentinvention can be employed to provide a skin appearance and/or feelbenefit. Quantities of the present compositions that are typicallyapplied per application are, in mg composition/cm² skin, from about 0.01mg/cm² to about 10 mg/cm². A particularly useful application amount isabout 1 mg/cm² to about 2 mg/cm².

[0231] Regulating keratinous tissue condition can be practiced byapplying a composition in the form of a skin lotion, cream, gel, foam,ointment, paste, emulsion, spray, conditioner, tonic, cosmetic,lipstick, foundation, nail polish, after-shave, or the like that ispreferably intended to be left on the skin or other keratin structurefor some esthetic, prophylactic, therapeutic or other benefit (i.e., a“leave-on” composition). After applying the composition to the skin, itcan be left on the skin for a period of at least about 15 minutes, or atleast about 30 minutes, or at least about 1 hour, or for at leastseveral hours, for example, up to about 12 hours.

[0232] Any part of the external portion of the face, hair, and/or nailscan be treated, e.g., face, lips, under-eye area; eyelids, scalp, neck,torso, arms, hands, legs, feet, fingernails, toenails, scalp hair,eyelashes, eyebrows, etc. The composition can be applied with thefingers or with an implement or device (e.g., pad, cotton ball,applicator pen, spray applicator, and the like).

[0233] Another approach to ensure a continuous exposure of the skin toat least a minimum level of the beneficial compositions of the inventionis to apply the composition in a patch, for example, to selected tissuessuch as the face. Such an approach is particularly useful for problemskin areas needing more intensive treatment (e.g., a wound, facial crowsfeet area, frown lines, under eye area, and the like). The patch can beocclusive, semi-occlusive or non-occlusive and can be adhesive ornon-adhesive. The composition can be contained within the patch or beapplied to the skin prior to application of the patch. The patch canalso include additional compounds such as chemical initiators forexothermic reactions such as those described in U.S. Pat. Nos.5,821,250, 5,981,547, and 5,972,957 to Wu, et al. The patch ispreferably left on the skin for a period of at least about 5 minutes, orat least about 15 minutes, or at least about 30 minutes, or at leastabout 1 hour, or at night as a form of night therapy.

[0234] The following examples are intended to further illustrate certainaspects of the invention but are not intended to be limiting thereof.

EXAMPLE 1 Stimulation of Fibroblast Growth

[0235] This Example provides data showing the cell proliferating effectof acetylsalicylic acid and salicylic acid on human skin fibroblasts.

[0236] Materials and Methods

[0237] A human skin fibroblast cell line (Clonetics, Walkersville, Md.,normal human dermal fibroblasts, neonatal) was tested to ascertainwhether exposure to acetylsalicylic acid and salicylic acid of theinvention would stimulate cellular proliferation. The proliferativeresponse of the human skin fibroblast cell line to acetylsalicylic acidand salicylic acid was measured in a 96-well assay system usingserum-free medium as a control. Both compounds were tested at threeconcentrations, 1×10⁻⁴ M, 1×10⁻⁵ M and 1×10⁻⁶ M. Cells (2×10³) wereseeded into a 96 well in 100 μl of Dulbecco's Modified Eagle's Medium(DMEM, Sigma Chemical Co., St. Louis, Mo.) containing 10% fetal bovineserum (FBS, Sigma Chemical Co., St. Louis, Mo.). The plate was incubatedfor 24 hours at 37° C. in a humidified 5% CO₂ atmosphere. Afterincubation, the medium was aspirated and the wells were rinsed twicewith 100 μl of serum-free DMEM. The final rinse was aspirated and 100 μlof the 1×10⁻⁴ M, 1×10⁻⁵ M or 1×10⁻⁶ M solutions of acetylsalicylic acidand salicylic acid each were added to 5 wells. In addition, 100 μl ofvehicle (serum-free DMEM) was added to 5 wells as control. All wellswere incubated for 28 hours at 37° C. in a humidified, 5% CO₂atmosphere. After incubation, 20 μl of Cell Titer 96 Aqueous OneSolution (Promega, Madison, Wis.) was added to all wells. The plate wasswirled gently and placed back in the incubator for 45 minutes andspectrophotometric absorbance was read at 490 nm.

[0238] Statistical analyses of data were performed using one-way ANOVA.P<0.05 is considered significant, while P<0.0001 is considered extremelysignificant and P<0.001 is considered as very very significant.

Results

[0239] Table 1 and FIG. 1 illustrate the cell proliferating effect ofacetylsalicylic acid on human skin fibroblasts, where the concentrationof acetylsalicylic acid varied between 1×10⁻⁴ M (designated ASA4),1×10⁻⁵ M (designated ASA5) and 1×10⁻⁶ M (designated ASA6). TABLE 1Effect of Acetylsalicylic acid on Cell Proliferation Number Standard ofStandard Error of Group Points Mean Deviation Mean Median Control 50.3784 0.009317 0.004167 0.3800 ASA4 5 0.5406 0.03051 0.01364 0.5290ASA5 5 0.6276 0.03310 0.01480 0.6270 ASA6 5 0.6590 0.04973 0.022240.6590

[0240] The data provided in Table 1 and FIG. 1 indicate thatacetylsalicylic acid has strong cell proliferating activity at allconcentrations (P<0.001 at all concentrations). The effect on cellproliferation was dose dependent, with a greater effect at lower, ratherthan higher, concentrations.

[0241] Table 2 and FIG. 2 illustrate the cell proliferating effect ofsalicylic acid on human skin fibroblasts, where the concentration ofsalicylic acid varied between 1×10⁻⁴ M (designated SA4), 1×10⁻⁵ M(designated SA5) and 1×10⁻⁶ M (designated SA6). TABLE 2 Effect ofSalicylic acid on Cell Proliferation Number Standard of Standard Errorof Group Points Mean Deviation Mean Median Control 5 0.3368 0.012010.005370 0.3310 SA4 5 0.3916 0.03116 0.01393 0.4050 SA5 5 0.5004 0.025320.01133 0.5110 SA6 5 0.4668 0.01599 0.007151 0.4690

[0242] The data provided in Table 2 and FIG. 2 indicates that salicylicacid has strong cell proliferating activity especially at the lowerconcentrations (P<0.001 at concentrations of 10⁻⁵ and 10⁻⁶ M). Theeffect on cell proliferation was dose dependent manner with a greatereffect at lower, rather than higher, concentrations.

EXAMPLE 2 Stimulation of Keratinocyte Growth

[0243] This Example provides data showing the cell proliferating effectof jasmonic acid, zeatin and gibberellic acid on human skinkeratinocytes.

[0244] Materials and Methods

[0245] A human skin keratinocyte cell line from Clonetics (Walkersville,Md., normal human epidermal keratinocytes, neonatal, catalog numbercc-2503) was exposed to the jasmonic acid, zeatin and gibberellic acid(from Sigma Chemical Co.) to determine their effect on proliferation ofkeratinocytes. The proliferative response of these human skinkeratinocytes to the test compounds was measured in a 96-well assaysystem using keratinocyte basal medium (KBM, Clonetics, catalog numberCC-3103) as a control. All compounds were tested at threeconcentrations, 1×10⁻⁴ M, 1×10⁻⁵ M and 1×10⁻⁶ M. Cells were seeded intoa 96 well plate at a concentration of 2×10³ cells in 100 μl of KBM. Theplate was incubated for 24 hours at 37° C. in a humidified 5% CO₂atmosphere. After incubation, 100 μl of the 1×10⁻⁴ M, 1×10⁻⁵ M or 1×10⁻⁶M solutions of jasmonic acid, zeatin and gibberellic acid were added to5 wells each. In addition, 100 μl of vehicle KBM was added to 5 wells ascontrol. The plate was incubated for 48 hours at 37° C. in a humidified,5% CO₂ atmosphere. After incubation, 20 μl of Cell Titer 96 Aqueous OneSolution (Promega, Madison, Wis.) was added to all wells. The plate wasswirled gently and placed back in the incubator for 3 hours. Thespectrophotometric absorbance of each well was read at 490 nm.

[0246] Statistical analyses of data were performed using one-way ANOVA.P<0.05 is considered significant, while P<0.0001 is considered extremelysignificant and P<0.001 is considered as very, very significant.

Results

[0247] Table 3 and FIG. 3 illustrate the cell proliferating effect ofjasmonic acid on human keratinocytes, where the concentration ofjasmonic acid varied between 1×10⁻⁴ M (designated JA4), 1×10⁻⁵ M(designated JA5) and 1×10⁻⁶ M (designated JA6). TABLE 3 Effect ofJasmonic Acid on Cell Proliferation Number Standard of Standard Error ofGroup Points Mean Deviation Mean Median JA4 5 0.5732 0.02720 0.012160.5860 JA5 5 0.7628 0.03805 0.01702 0.7750 JA6 5 0.7734 0.06328 0.028300.7970 Control 5 0.5094 0.07334 0.03280 0.5050

[0248] The data provided in Table 3 and FIG. 3 indicates that jasmonicacid has strong cell proliferating activity especially at the lowerconcentrations (P<0.001 at concentrations of 10⁻⁵ and 10⁻⁶ M). Theeffect on cell proliferation was greater at lower, rather than higher,concentrations.

[0249] Table 4 and FIG. 4 illustrate the cell proliferating effect oft-zeatin on human keratinocytes, where the concentration of t-zeatinvaried between 1×10⁻⁴ M (designated ZA4), 1×10⁻⁵ M (designated ZAS) and1×10⁻⁶ M (designated ZA6). TABLE 4 Effect of t-Zeatin on CellProliferation Number Standard of Standard Error of Group Points MeanDeviation Mean Median ZA4 5 0.7726 0.03785 0.01693 0.7880 ZA5 5 0.87160.03179 0.01421 0.8790 ZA6 5 0.9770 0.05141 0.02299 0.9830 Control 50.5336 0.07444 0.03329 0.5650

[0250] The data provided in Table 4 and FIG. 4 indicate that t-zeatinhas strong cell proliferating activity at all concentrations (P<0.001 atall concentrations). The effect on cell proliferation was dose dependentmanner with a greater effect at lower, rather than higher,concentrations.

[0251] Table 5 and FIG. 5 illustrate the cell proliferating effect ofgibberellic acid on human keratinocytes, where the concentration ofgibberellic acid varied between 1×10⁻⁴ M (designated GA4), 1×10⁻⁵ M(designated GAS) and 1×10⁻⁶ M (designated GA6). TABLE 5 Effect ofGibberellic acid on Cell Proliferation Number Standard of Standard Errorof Group Points Mean Deviation Mean Median +TL,1GA4 5 0.6512 0.012560.005616 0.6510 GA5 5 0.8184 0.04813 0.02152 0.8210 GA6 5 0.7854 0.047430.02121 0.7900 Control 5 0.5308 0.06833 0.03056 0.5220

[0252] The data provided in Table 5 and FIG. 5 indicate that giberrellicacid has strong cell proliferating activity at all concentrations(P<0.001 at all concentrations). The effect on cell proliferation wasgreater at lower, rather than higher, concentrations.

EXAMPLE 3 Stimulation of Collagen Production

[0253] The example provides data showing the effect of jasmonic acid,zeatin and gibberellic acid on collagen production.

[0254] The stimulation response of jasmonic acid, zeatin and gibberellicacid on collagen production in the human skin fibroblast cell line(Clonetics, Walkersville, Md., normal human dermal fibroblasts,neonatal, catalog number CC-2509) was measured using Takara BiomedicalsEIA assay kit (TAK MK101) sold by Panvera (Madison, Wis.). The cellswere first grown in a 96-well assay system using Dulbecco's ModifiedEagle's Medium (DMEM) with 10% fetal bovine serum (FBS) both purchasedfrom Sigma Chemical Co, St. Louis, Mo. Serum-free DMEM was used as acontrol. All compounds were tested at three concentrations 1×10⁻⁴ M,1×10⁻⁵ M and 1×10⁻⁶ M. Cells were seeded into a 96 well plate at aconcentration of 5×10³ cells in 100 μl of DMEM containing 10% fetalbovine serum (FBS, Sigma Chemical Co., St. Louis, Mo.). Plate wasincubated for 24 hours at 37° C. in a humidified 5% CO₂ atmosphere.After incubation, the medium was aspirated and the wells were rinsedtwice with 100 μl of serum-free DMEM. The final rinse was aspirated and100 μl of the 1×10⁻⁴ M, 1×10⁻⁵ M or 1×10⁻⁶ M solutions of the testcompounds were added to the wells (n=2 for each concentration). Inaddition, 100 μl of vehicle (serum-free DMEM) was added to 4 wells ascontrol. The plate was incubated for 48 hours at 37° C. in a humidified,5% CO₂ atmosphere.

[0255] The assay was done by using the recommended 20 ul of thesupernatant from each well of the 96-well plate. Standard buffer andstop solutions were freshly prepared before running the assay. 100 ul ofantibody-POD conjugate solution (supplied with the kit) was added intothe wells using pre antibody coated 96 well plate (supplied with thekit). 20 ul of standard and test solutions (from the other 96-well platecontaining fibroblasts) were added to appropriate wells. Plate was mixedgently, sealed and incubated for three hrs. at 37° C.

[0256] After incubation each well was washed carefully four times withPBS buffer (400 ul). All the wells were completely emptied at the end ofwashing from any liquid.

[0257] 100 ul of substrate solution (hydrogen peroxide andtetramethylbenzidine in a buffer solution, supplied with the kit) wasadded to each well and the plate was incubated for 15 minutes. At thispoint 100 ul of stop solution (freshly prepared 1N H₂SO₄) was added toeach well in the same order as substrate. The plate was gently mixed andabsorbance was read at 450 nm.

[0258] Statistical analyses of data were performed using one-way ANOVA.P<0.05 is considered significant, while P<0.0001 is considered extremelysignificant and P<0.001 is considered as very, very significant.

Results

[0259] Table 6 and FIG. 6 illustrate the collagen production of humanfibroblast cells exposed to varying concentrations of jasmonic acid,gibberellic acid or t-zeatin. The concentration of jasmonic acid variedbetween 1×10⁻⁴ M (designated JA4), 1×10⁻⁵ M (designated JA5) and 1×10⁻⁶M (designated JA6). The concentration of gibberellic acid varied between1×10⁻⁴ M (designated GA4), 1×10⁻⁵ M (designated GA5) and 1×10⁻⁶ M(designated GA6). The concentration of zeatin varied between 1×10⁻⁴ M(designated ZA4), 1×10⁻⁵ M (designated ZA5) and 1×10⁻⁶ M (designatedZA6). TABLE 6 Effect of Jasmonic Acid, Gibberellic Acid and Zeatin onCollagen Production by Human Fibroblasts Number Standard of StandardError of Group Points Mean Deviation Mean Median JA4 2 0.8040 0.0098990.007000 0.8040 JA5 2 0.7895 0.006364 0.004500 0.7895 JA6 2 0.74750.007778 0.005500 0.7475 GA4 2 0.7780 0.01838 0.01300 0.7780 GA5 20.8365 0.04172 0.02950 0.8365 GA6 2 0.8630 0.008485 0.006000 0.8630 ZA42 0.7665 0.02758 0.01950 0.7665 ZA5 2 0.8115 0.0007071 0.0005000 0.8115ZA6 2 0.7715 0.006364 0.004500 0.7715 CNA5 4 0.7483 0.006238 0.0031190.7495 Control 4 0.7388 0.01431 0.007157 0.7330

[0260] The data provided in Table 6 and FIG. 6 indicate that jasmonicacid, giberrellic acid and zeatin can all stimulate collagen productionin human fibroblasts. The effect of gibberellic acid on collagenproduction was more profound, particularly at lower, rather than higher,concentrations. However, jasmonic acid strongly stimulated collagenproduction at higher concentrations and zeatin strongly stimulatedcollagen production at medium concentrations (see FIG. 6).

[0261] All publications and patents are incorporated by referenceherein, as though individually incorporated by reference. The inventionis not limited to the exact details shown and described, for it shouldbe understood that many variations and modifications may be made whileremaining within the spirit and scope of the invention defined by thestatements.

1 3 1 4 PRT Artificial Sequence A synthesized tetrapeptide 1 Arg Ser ArgLys 1 2 5 PRT Artificial Sequence A synthesized pentapeptide 2 Lys ThrThr Lys Ser 1 5 3 5 PRT Artificial Sequence A synthesized pentapeptidederivative having the sequence palmitoyl-Lys- Thr-Thr-Lys-Ser 3 Lys ThrThr Lys Ser 1

What is claimed:
 1. A composition comprising a dermatologicallyacceptable carrier and an effective amount of a hydroxy acid compound,wherein the composition can stimulate cellular proliferation ofmammalian fibroblasts or keratinocytes, and wherein the hydroxy acidcompound is a compound of formula I:

wherein: R1 is COOR, or —(CH₂)n-OX; R is H, or alkyl; n is an integer ofabout 1 to about 20; X is H, or 1 to 6 sugar residues; R2 is COOR,—(CH₂)n-OX, OCO-alkyl, OY; and Y is H, alkyl, or 1 to 6 sugar residues.2. The composition of claim 1, wherein hydroxy acid is salicylic acid,lactic acid, glycolic acid, or acetylsalicylic acid.
 3. The compositionof claim 1, wherein the composition further comprises an effectiveamount of a jasmonic acid compound, a gibberellic acid compound or azeatin compound.
 4. The composition of claim 3, wherein the jasmonicacid compound is a compound of any one of formulae VI-IX:

wherein: R3 is alkyl; R4 is COOR, or —(CH₂)n-OX, where n is an integerof from 1 to 20; R is H, or alkyl; X is H, or 1 to 6 sugar residues; andY is H, alkyl, or 1 to 6 sugar residues.
 5. The composition of claim 3,wherein the jasmonic acid compound is jasmonic acid, hydroxyjasmonicacid, dihydrojasmonic acid, or dihydro-hydroxy jasmonic acid.
 6. Thecomposition of claim 3, wherein the zeatin compound is a compound offormula X:

wherein: R5 is H, 3-hydroxymethl-3-methylallyl, alkyl, —(CH₂)n-CH₃, orOZ; Z is H, 1 to 6 sugar residues, or —(CH₂)n-furan; R6 is H,3-hydroxymethl-3-methylallyl, alkyl, —(CH₂)n-CH₃, or OZ; and n is aninteger of 1 to
 20. 7. The composition of claim 3, wherein the zeatincompound is zeatin.
 8. The composition according to claim 1, wherein theeffective amount of the hydroxy acid compound is a concentration ofabout 0.001 micromolar to about 10 millimolar.
 9. The compositionaccording to claim 1, wherein the effective amount of the hydroxy acidcompound is a concentration of about 1 micromolar to about 5 millimolar.10. The composition according to claim 1, further comprising one or moreadditional ingredients.
 11. The composition of claim 10, wherein theadditional ingredient is a desquamation compound, an anti-acne compound,an anti-wrinkle compound, a vitamin B₃ compound, a vitamin E compound, aretinoid compound, a hydroxy acid compound, an anti-oxidant compound, aradical scavenger, a chelating agent, a flavonoid compound, ananti-inflammatory agent, an anti-cellulite agent, a topical anesthetic,a tanning compound, a skin lightening agent, a skin healing compound, anantimicrobial compound, an antifungal compound, a sunscreen compound, aparticulate material, a moisturizer, or a thickening agent.
 12. Thecomposition of claim 11, wherein the vitamin E compound is tocopherol,tocopherol acetate, a tocopherol ester or a mixture thereof.
 13. Thecomposition of claim 11, wherein the retinoid compound is a retinol,retinal, retinol ester, retinyl propionate, retinoic acid, retinylpalmitate, or a mixture thereof.
 14. The composition of claim 11,wherein the particulate material is mica, mica treated with bariumsulfate and TiO₂, silica, nylon, polyethylene, talc, styrene,polypropylene, ethylene/acrylic acid copolymer, sericite, aluminumoxide, silicone resin, barium sulfate, titanium dioxide, iron oxide,bismuth oxychloride, calcium carbonate, cellulose acetate, polymethylmethacrylate, or a mixture thereof.
 15. The composition of claim 11,wherein the sunscreen is a metallic oxide selected from the groupconsisting of titanium dioxide having an average primary particle sizeof from about 15 nm to about 100 nm, zinc oxide having an averageprimary particle size of from about 15 nm to about 150 nm, zirconiumoxide having an average primary particle size of from about 15 nm toabout 150 nm, iron oxide having an average primary particle size of fromabout 15 nm to about 500 nm, or a mixture thereof.
 16. The compositionof claim 11, wherein the sunscreen is octylmethoxycinnamate, octylsalicylate, terephthalyidene dicamphor sulfonic acid, avobenzone,octocrylene, or a mixture thereof.
 17. A composition comprising adermatologically acceptable carrier and an effective amount of ajasmonic acid compound, a gibberellic acid compound or a zeatincompound; wherein the composition can stimulate cellular proliferationof mammalian fibroblasts or keratinocytes.
 18. The composition of claim17, wherein the jasmonic acid compound is a compound of any one offormulae VI-IX:

wherein: R3 is alkyl; R4 is COOR, or —(CH₂)n-OX, where n is an integerof from 1 to 20; R is H, or alkyl; X is H, or 1 to 6 sugar residues; andY is H, alkyl, or 1 to 6 sugar residues.
 19. The composition of claim17, wherein the jasmonic acid compound is jasmonic acid, hydroxyjasmonicacid, dihydrojasmonic acid, or dihydro-hydroxy jasmonic acid.
 20. Thecomposition of claim 17, wherein the zeatin compound is a compound offormula X:

wherein: R5 is H, 3-hydroxymethl-3-methylallyl, alkyl, —(CH₂)n-CH₃, orOZ; Z is H, 1 to 6 sugar residues, or —(CH₂)n-furan; R6 is H,3-hydroxymethl-3-methylallyl, alkyl, —(CH₂)n-CH₃, or OZ; and n is aninteger of 1 to
 20. 21. The composition of claim 17, wherein the zeatincompound is zeatin.
 22. The composition of claim 17, wherein thecomposition further comprises a hydroxy acid compound of formula I:

wherein: R1 is COOR, or —(CH₂)n-OX; R is H, or alkyl; n is an integer ofabout 1 to about 20; X is H, or 1 to 6 sugar residues; R2 is COOR,—(CH₂)n-OX, OCO-alkyl, OY; and Y is H, alkyl, or 1 to 6 sugar residues.23. The composition of claim 17, wherein the effective amount of thejasmonic acid compound, gibberellic acid compound or zeatin compound isa concentration of about 0.001 micromolar to about 10 millimolar.
 24. Acomposition comprising a dermatologically acceptable carrier and aneffective amount of a jasmonic acid compound, a gibberellic acidcompound or a zeatin compound; wherein the composition can stimulatecollagen production in mammalian fibroblasts.
 25. The composition ofclaim 24, wherein the jasmonic acid compound is a compound of any one offormulae VI-IX:

wherein: R3 is alkyl; R4 is COOR, or —(CH₂)n-OX, where n is an integerof from 1 to 20; R is H, or alkyl; X is H, or 1 to 6 sugar residues; andY is H, alkyl, or 1 to 6 sugar residues.
 26. The composition of claim24, wherein the jasmonic acid compound is jasmonic acid, hydroxyjasmonicacid, dihydrojasmonic acid, or dihydro-hydroxyjasmonic acid.
 27. Thecomposition of claim 24, wherein the zeatin compound is a compound offormula X:

wherein: R5 is H, 3-hydroxymethl-3-methylallyl, alkyl, —(CH₂)n-CH₃, orOZ; Z is H, 1 to 6 sugar residues, or —(CH₂)n-furan; R6 is H,3-hydroxymethl-3-methylallyl, alkyl, —(CH₂)n-CH₃, or OZ; and n is aninteger of 1 to
 20. 28. The composition of claim 24, wherein the zeatincompound is zeatin.
 29. The composition of claim 24, wherein thecomposition further comprises hydroxy acid compound of formula I:

wherein: R1 is COOR, or —(CH₂)n-OX; R is H, or alkyl; n is an integer ofabout 1 to about 20; X is H, or 1 to 6 sugar residues; R2 is COOR,—(CH₂)n-OX, OCO-alkyl, OY; and Y is H, alkyl, or 1 to 6 sugar residues.30. The composition of claim 24, wherein the effective amount of thejasmonic acid compound, gibberellic acid compound or zeatin compound isa concentration of about 0.001 micromolar to about 10 millimolar. 31.The composition of claim 24, wherein the effective amount of thejasmonic acid compound, gibberellic acid compound or zeatin compound isa concentration of about 1 micromolar to about 5 millimolar.
 32. Thecomposition of claim 24, further comprising one or more additionalingredients.
 33. The composition of claim 32, wherein the additionalingredient is a desquamation compound, an anti-acne compound, ananti-wrinkle compound, a vitamin B₃ compound, a vitamin E compound, aretinoid compound, a hydroxy acid compound, an anti-oxidant compound, aradical scavenger, a chelating agent, a flavonoid compound, ananti-inflammatory agent, an anti-cellulite agent, a topical anesthetic,a tanning compound, a skin lightening agent, a skin healing compound, anantimicrobial compound, an antifungal compound, a sunscreen compound, aparticulate material, a moisturizer, or a thickening agent.
 34. Thecomposition of claim 33, wherein the vitamin E compound is tocopherol,tocopherol acetate, a tocopherol ester or a mixture thereof.
 35. Thecomposition of claim 33, wherein the retinoid compound is a retinol,retinal, retinol ester, retinyl propionate, retinoic acid, retinylpalmitate, or a mixture thereof.
 36. The composition of claim 33,wherein the particulate material is mica, mica treated with bariumsulfate and TiO₂, silica, nylon, polyethylene, talc, styrene,polypropylene, ethylene/acrylic acid copolymer, sericite, aluminumoxide, silicone resin, barium sulfate, titanium dioxide, iron oxide,bismuth oxychloride, calcium carbonate, cellulose acetate, polymethylmethacrylate, or a mixture thereof.
 37. The composition of claim 33,wherein the sunscreen is a metallic oxide selected from the groupconsisting of titanium dioxide having an average primary particle sizeof from about 15 nm to about 100 nm, zinc oxide having an averageprimary particle size of from about 15 nm to about 150 nm, zirconiumoxide having an average primary particle size of from about 15 nm toabout 150 nm, iron oxide having an average primary particle size of fromabout 15 nm to about 500 nm, or a mixture thereof.
 38. The compositionof claim 33, wherein the sunscreen is octylmethoxycinnamate, octylsalicylate, terephthalyidene dicamphor sulfonic acid, avobenzone,octocrylene, or a mixture thereof.
 39. A skin care compositioncomprising: a) from about 0.01% to about 30%, by weight of thecomposition, of jasmonic acid, gibberellic acid or zeatin; b) from about0.01% to about 30%, by weight of the composition, of acetylsalicylicacid or salicylic acid; and c) a dermatologically acceptable carrier.40. An orally acceptable formulation for the treatment of gum tissuescomprising an acceptable carrier and an effective amount of a jasmonicacid compound, a gibberellic acid compound or a zeatin compound; whereinthe formulation can stimulate cellular proliferation of mammalianfibroblasts or keratinocytes.
 41. The formulation of claim 40, whereinthe formulation can stimulate collagen production in mammalianfibroblasts.
 42. The formulation of claim 40, wherein the jasmonic acidcompound is a compound of any one of formulae VI-IX:

wherein: R3 is alkyl; R4 is COOR, or —(CH₂)n-OX, where n is an integerof from 1 to 20; R is H, or alkyl; X is H, or 1 to 6 sugar residues; andY is H, alkyl, or 1 to 6 sugar residues.
 43. The formulation of claim40, wherein the jasmonic acid compound is jasmonic acid, hydroxyjasmonicacid, dihydrojasmonic acid, or dihydro-hydroxyasmonic acid.
 44. Theformulation of claim 40, wherein the zeatin compound is a compound offormula X:

wherein: R5 is H, 3-hydroxymethl-3-methylallyl, alkyl, —(CH₂)n-CH₃, orOZ; Z is H, 1 to 6 sugar residues, or —(CH₂)n-furan; R6 is H,3-hydroxymethl-3-methylallyl, alkyl, —(CH₂)n-CH₃, or OZ; and n is aninteger of 1 to
 20. 45. The formulation of claim 40, wherein the zeatincompound is zeatin.
 46. The formulation of claim 40, wherein thecomposition further comprises a hydroxy acid compound of formula I:

wherein: R1 is COOR, or —(CH₂)n-OX; R is H, or alkyl; n is an integer ofabout 1 to about 20; X is H, or 1 to 6 sugar residues; R2 is COOR,—(CH₂)n-OX, OCO-alkyl, OY; and Y is H, alkyl, or 1 to 6 sugar residues.47. The formulation of claim 40, wherein the effective amount of thejasmonic acid compound, gibberellic acid compound or zeatin compound isa concentration of about 0.001 micromolar to about 10 millimolar.
 48. Amethod of stimulating growth of fibroblasts or keratinocytes comprisingadministering to the fibroblasts or the keratinocytes a safe andeffective amount of a composition comprising a dermatologicallyacceptable carrier and an effective amount of a hydroxy acid compound,wherein the composition can stimulate cellular proliferation ofmammalian fibroblasts or keratinocytes, and wherein the hydroxy acidcompound comprises a compound of formula I:

wherein: R1 is COOR, or —(CH₂)n-OX; R is H, or alkyl; n is an integer ofabout 1 to about 20; X is H, or 1 to 6 sugar residues; R2 is COOR,—(CH₂)n-OX, OCO-alkyl, OY; and Y is H, alkyl, or 1 to 6 sugar residues.49. A method of stimulating growth of fibroblasts or keratinocytescomprising administering to the fibroblasts or the keratinocytes a safeand effective amount of a composition comprising a dermatologicallyacceptable carrier and an effective amount of a jasmonic acid compound,a gibberellic acid compound or a zeatin compound; wherein thecomposition can stimulate cellular proliferation of mammalianfibroblasts or keratinocytes.
 50. A method of stimulating collagenproduction in mammalian fibroblasts comprising administering to thefibroblasts a safe and effective amount of a composition comprising adermatologically acceptable carrier and an effective amount of ajasmonic acid compound, a gibberellic acid compound or a zeatincompound; wherein the composition can stimulate collagen production inmammalian fibroblasts.